Inhibition of antibacterial activity of himastatin, a new antitumor antibiotic from Streptomyces hygroscopicus, by fatty sodium salts.

Himastatin, a cyclohexadepsipeptide antibiotic, had in vivo antitumor activity against localized P388 leukemia and B16 melanoma but had no distal site antitumor activity. An in vitro Bacillus subtilis well-agar diffusion assay was employed to test the hypothesis that himastatin was enzymatically inactivated. The activity of himastatin against B. subtilis was inhibited when himastatin was mixed with mouse liver S9 fraction and microsomes. However, subsequent investigations demonstrated that the markedly decreased antibacterial activity was not enzymatic in nature but was related to the presence of certain fatty salts. Saturated fatty sodium salts with a carbon chain number of 8 or more reduced the antimicrobial activity of himastatin 50 to 100 times. If antibiotics such as ampicillin, bacitracin, chloramphenicol, and tunicamycin were used in place of himastatin, no meaningful reduction in antibacterial activity occurred. However, the antibacterial activity of the membrane-active peptide antibiotic polymyxin B, but not that of polymyxin E (colistin), was reduced in a manner similar to that of himastatin. Importantly, the activity of himastatin against HCT-116 adenocarcinoma cells in soft agar was markedly reduced in the presence of sodium palmitate as the reference fatty salt. The data indicate that himastatin may be trapped in micelles in vitro. It may be speculated that the lack of distal site antitumor activity resulted from similar complex formation between himastatin and lipids in vivo. The results also suggest that the cytotoxic and antimicrobial effects of himastatin may result from interactions with the cell membrane.

Keyword: colon cancer

Fatty composition and anticancer activity in carcinoma cell lines of Prunus dulcis seed oil.

Almond oil is used in traditional and complementary therapies for its numerous health benefits due to high unsaturated fatty acids content.This study investigated the composition and in vitro anticancer activity of almond oil from Northern Cyprus and compared with almond oil from Turkey.Almond oil from Northern Cyprus was obtained by supercritical CO extraction and analyzed by GC-MS. Almond oil of Turkey was provided from Turkish pharmacies. Different concentrations of almond oils were incubated for 24 and 48\u2009h with Colo-320 and Colo-741 cells. Cell growth and cytotoxicity were measured by MTT assays. Anticancer and antiprolifetarive activities of almond oils were investigated by immunocytochemistry using antibodies directed against to BMP-2, β-catenin, Ki-67, LGR-5 and Jagged 1.Oleic (77.8%; 75.3%), linoleic (13.5%; 15.8%), (7.4%; 6.3%), were determined as the major compounds of almond oil from Northern Cyprus and Turkey, respectively. In the MTT assay, both almond oils were found to be active against Colo-320 and Colo-741 cells with 1:1 dilution for both 24\u2009h and 48\u2009h. As a result of immunohistochemical staining, while both almond oils exhibited significant antiproliferative and anticancer activity, these activities were more similar in Colo-320 cells which were treated with Northern Cyprus almond oil.Almond oil from Northern Cyprus and Turkey may have anticancer and antiproliferative effects on cells through molecular signalling pathways and, thus, they could be potential novel therapeutic agents.

Keyword: colon cancer

5'-O-Palmitoyl- and 3',5'-O-dipalmitoyl-5-fluoro-2'-deoxyuridine--novel lipophilic analogues of 5'-fluoro-2'-deoxyuridine: synthesis, incorporation into liposomes and preliminary biological results.

5'-O-palmitoyl- and 3',5'-O-dipalmitoyl-5-fluoro-2'-deoxyuridine were prepared by the reaction of 5-fluoro-2'-deoxyuridine in dimethylacetamide with chloride. The incorporation of the synthesized prodrugs into liposomes composed of egg phosphatidylcholine/stearylamine/cholesterol/alpha-tocopherol at a molar ratio of 10:1:2:0.05 was nearly quantitative; homogeneous bilayer vesicles (75 nm diameter) were obtained. Preliminary tolerance studies revealed that the prodrug-liposome preparations are about 20-60 times more toxic than the parent drug. The prodrugs incorporated into liposomes were 10 to 30 times more active against murine 38 carcinoma compared to the free drug. In comparison to the administration of the prodrugs in peanut oil the liposomal preparations seem to exert improved effects and represent a valuable drug delivery system for parenteral applications.

Keyword: colon cancer

Myristoylation of heparan sulfate proteoglycan and proteins occurs post-translationally in human carcinoma cells.

We have recently shown that the heparan sulfate proteoglycan of human carcinoma cells is acylated with both myristate and palmitate, two long-chain saturated fatty acids. In this study we show that cycloheximide did not significantly inhibit the incorporation of myristic into either proteoglycan or total protein pool. This lack of inhibition occurred under a condition in which protein synthesis was inhibited greater than 90%. Cycloheximide, on the other hand, did not affect the incorporation of [3H]myristic into fatty nor the intracellular interconversion of myristate to palmitate. Characterization of fatty acyl moiety in the proteoglycan and protein by reverse-phase HPLC revealed that approximately 60% of the covalently bound fatty acids was myristate and the remaining 40% was palmitate. These results indicate that in human carcinoma cells myristoylation of heparan sulfate proteoglycan and proteins occurs post-translationally, presumably in the Golgi complex.

Keyword: colon cancer

Aldo-keto reductase family 1 member B10 promotes cell survival by regulating lipid synthesis and eliminating carbonyls.

Aldo-keto reductase family 1 member B10 (AKR1B10) is primarily expressed in the normal human and small intestine but overexpressed in liver and lung . Our previous studies have shown that AKR1B10 mediates the ubiquitin-dependent degradation of acetyl-CoA carboxylase-alpha. In this study, we demonstrate that AKR1B10 is critical to cell survival. In human carcinoma cells (HCT-8) and lung carcinoma cells (NCI-H460), small-interfering RNA-induced AKR1B10 silencing resulted in caspase-3-mediated apoptosis. In these cells, the total and subspecies of cellular lipids, particularly of phospholipids, were decreased by more than 50%, concomitant with 2-3-fold increase in reactive oxygen species, mitochondrial cytochrome c efflux, and caspase-3 cleavage. AKR1B10 silencing also increased the levels of alpha,beta-unsaturated carbonyls, leading to the 2-3-fold increase of cellular lipid peroxides. Supplementing the HCT-8 cells with (80 mum), the end product of fatty synthesis, partially rescued the apoptosis induced by AKR1B10 silencing, whereas exposing the HCT-8 cells to epalrestat, an AKR1B10 inhibitor, led to more than 2-fold elevation of the intracellular lipid peroxides, resulting in apoptosis. These data suggest that AKR1B10 affects cell survival through modulating lipid synthesis, mitochondrial function, and oxidative status, as well as carbonyl levels, being an important cell survival protein.

Keyword: colon cancer

Human carbonyl reductase 1 participating in intestinal first-pass drug metabolism is inhibited by fatty acids and acyl-CoAs.

Human carbonyl reductase 1 (CBR1), a member of the short-chain dehydrogenase/reductase (SDR) superfamily, reduces a variety of carbonyl compounds including endogenous isatin, prostaglandin E and 4-oxo-2-nonenal. It is also a major non-cytochrome P450 enzyme in the phase I metabolism of carbonyl-containing drugs, and is highly expressed in the intestine. In this study, we found that long-chain fatty acids and their CoA ester derivatives inhibit CBR1. Among saturated fatty acids, myristic, and stearic acids were inhibitory, and stearic was the most potent (IC 9µM). Unsaturated fatty acids (oleic, elaidic, γ-linolenic and docosahexaenoic acids) and acyl-CoAs (palmitoyl-, stearoyl- and oleoyl-CoAs) were more potent inhibitors (IC 1.0-2.5µM), and showed high inhibitory selectivity to CBR1 over its isozyme CBR3 and other SDR superfamily enzymes (DCXR and DHRS4) with CBR activity. The inhibition by these fatty acids and acyl-CoAs was competitive with respect to the substrate, showing the K values of 0.49-1.2µM. Site-directed mutagenesis of the substrate-binding residues of CBR1 suggested that the interactions between the fatty acyl chain and the enzyme's Met141 and Trp229 are important for the inhibitory selectivity. We also examined CBR1 inhibition by oleic in cellular levels: The fatty effectively inhibited CBR1-mediated 4-oxo-2-nonenal metabolism in DLD1 cells and increased sensitivity to doxorubicin in the drug-resistant gastric MKN45 cells that highly express CBR1. The results suggest a possible new food-drug interaction through inhibition of CBR1-mediated intestinal first-pass drug metabolism by dietary fatty acids.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: colon cancer

Characterization of posttranslational modifications of human A33 antigen, a novel palmitoylated surface glycoprotein of human gastrointestinal epithelium.

Monoclonal antibody (mAb) A33 recognizes a differentiation antigen (A33) expressed in normal human gastrointestinal epithelium and in 95% of human cancers. Murine mAb A33 shows specific targeting of in humans and a humanized A33 antibody is currently being evaluated in the clinic. The cDNA for the human A33 antigen has recently been cloned, and sequence comparison indicated that the A33 antigen is a novel human cell surface molecule of the immunoglobulin superfamily. Because mAb A33 recognizes a conformational epitope, only a partial characterization of the A33 antigen has been carried out to date. In this report we show that the A33 antigen is (I) N-glycosylated, containing approximately 8 K of N-linked carbohydrate and there is no evidence for O-glycosylation, sialylation or glycophosphatidylinositol, and (ii) S-acylated in vitro, incorporating [3H] linked through a hydroxylamine-sensitive thioester bond. The S-palmitoylation may be involved in regulating the internalization process initiated by binding of mAb A33 to cell surface A33 antigen.

Keyword: colon cancer

In vitro anti-inflammatory and anti-proliferative activity of sulfolipids from the red alga Porphyridium cruentum.

A sulfoglycolipidic fraction (SF) isolated from the red microalga Porphyridium cruentum was analyzed for fatty composition and assayed for ability to inhibit, in vitro, the generation of superoxide anion in primed leucocytes and the proliferation of a panel of human cell-lines. Results demonstrated that SF contained large amounts of (26.1%), arachidonic (C20: 4 omega-6, 36.8%), and eicopentaenoic (C20:5 omega-3, 16.6%) acids, and noticeable amounts of 16:1n-9 fatty (10.5%). It strongly inhibited both the production of superoxide anion generated by peritoneal leukocytes primed with phorbol myristate acetate (IC(50): 29.5 microg/mL), and the growth of human adenocarcinoma DLD-1 and to a lesser extent of human breast adenocarcinoma MCF-7, human prostate adenocarcinoma PC-3, and human malignant melanoma M4 Beu cell-lines, and therefore might have a chemopreventive or chemotherapeutic potential, or both. It was found markedly more cytotoxic than sulfoquinovosyldiacylglycerols from plant used as a standard (STD), due to a stronger ability to inhibit DNA alpha-polymerase (IC(50): 378 microg/mL, vs 1784 microg/mL for STD). After a 48-h continuous treatment, IC(50) values for growth inhibition were in the range of 20-46 microg/mL instead of 94 to >250 microg/mL for STD, and those for inhibition of metabolic activity were in the range of 34-87 microg/mL instead of >250 microg/mL for STD. The higher anti-proliferative effect was observed on adenocarcinoma DLD-1 cells, and the weaker effect was observed on breast adenocarcinoma MCF-7.

Keyword: colon cancer

Antimutagenicity of yogurt.

Yogurt is milk fermented by a mixture of two bacteria: Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus. Epidemiological studies have correlated a reduced risk of with yogurt consumption. Independent studies have established that yogurt and extracts thereof are antimutagenic. Although multiple explanations can account for yogurt's putative anticarcinogenicity, we are interested in testing the hypothesis that antimutagenic compounds produced during fermentation are responsible. We recently reported on the antimutagenicity of an acetone extract of yogurt against the experimental carcinogens N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 3.2'dimethyl-4-aminobiphenyl (DMAB) (Mutation Res. (1995) 334, 213-224). We are now aware that is an active ingredient against MNNG.

Keyword: colon cancer

Comparison of phospholipid profiles of primary adenocarcinoma in the lung and other organs.

Phospholipid profiles, particularly molecular structure of phosphatidylcholine, of human primary lung adenocarcinoma were compared with those of the histologically same type of carcinoma from other organs in order to search for a possibility that differentiates between primary and metastatic tumors in the lung. The saturated class, mainly containing at both positions, accounted for 20.8% of phosphatidylcholine in lung adenocarcinoma, whereas it accounted for only 6-10.7% in the adenocarcinoma of other organs. Adenocarcinoma of organs other than the lung had specific characteristics of unsaturated molecular classes of phosphatidylcholine for each organ. Stomach tumor had high contents of dienes and tetraenes with particularly high proportion of arachidonic at the 2-position. Breast tumor had a high content of monoenes, containing palmitoleic at the 2-position. Adenocarcinomas of rectum, and thyroid contained more dienes compared to lung adenocarcinoma.

Keyword: colon cancer

Fecal diglycerides as selective endogenous mitogens for premalignant and malignant human epithelial cells.

Diglycerides (DGs) have been found in fecal extracts at concentrations which induce mitogenesis of adenoma and some carcinoma cells but not normal cells in primary culture. DGs containing stearic, oleic, , and myristic side chains were found in fecal extracts from each of eight subjects. Synthetic 1,2-DGs, containing the fatty acids found in endogenous fecal DGs, induced mitogenesis in cultures of premalignant cells from each of 13 adenomas, covering all histological classes, and in cultures from two of four carcinomas. The potent adenoma mitogen, dimyristin, had no mitogenic activity on cultures of normal epithelial cells from seven different subjects. These results suggest DGs may act as endogenous mitogens in the development of human . The extent of adenoma mitogenesis was correlated with the chain length of the saturated R-groups: 16 greater than 14 greater than 12 greater than 10 greater than 8 much greater than 18. DGs with oleic residues, C18:1, were among the most active, while substitution of even one fatty residue with a stearic residue, C18:0, reduced or eliminated mitogenic activity. Dimyristin also induced enhanced levels of urokinase secretion from carcinoma cells, in parallel to the phorbol ester tumor promoter, 12-O-tetradecanoylphorbol-13-acetate. These results imply that DGs found in the induce a selective growth of benign tumors and some carcinomas, and may enhance the invasive capacity of carcinomas, while leaving normal cells unaffected.

Keyword: colon cancer

PKCζ Phosphorylates SIRT6 to Mediate Fatty β-Oxidation in Cells.

Protein kinase C (PKC) has critical roles in regulating lipid anabolism and catabolism. PKCζ, a member of atypical PKC family, has been reported to mediate glucose metabolism. However, whether and how PKCζ regulates tumor cells fatty β-oxidation are unknown. Here, we report that the phosphorylation of SIRT6 is significantly increased after (PA) treatment in cells. PKCζ can physically interact with SIRT6 in vitro and in vivo, and this interaction enhances following PA treatment. Further experiments show that PKCζ is the phosphorylase of SIRT6 and phosphorylates SIRT6 at threonine 294 residue to promote SIRT6 enrichment on chromatin. In the functional study, we find that the expression of ACSL1, CPT1, CACT, and HADHB, the genes related to fatty β-oxidation, increases after PA stimulation. We further confirm that PKCζ mediates the binding of SIRT6 specifically to the promoters of fatty β-oxidation-related genes and elicits the expression of these genes through SIRT6 phosphorylation. Our findings demonstrate the mechanism of PKCζ as a new phosphorylase of SIRT6 on maintaining tumor fatty β-oxidation and define the new role of PKCζ in lipid homeostasis.Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: colon cancer

The use of sesame oil and other vegetable oils in the inhibition of human growth in vitro.

Sesame contains large quantities of the essential polyunsaturated fatty (PUFA), linoleic , in the form triglycerides. The antineoplastic properties of many PUFAs such as linoleic and their metabolites are known. We tested the hypothesis that natural vegetable oils, such as sesame oil and its component linoleic , when added to human adenocarcinoma cells growing in tissue culture would inhibit their growth and that normal cells would not be similarly affected. Three human cell lines and one normal human cell line were exposed to the following: (1) pure linoleic ; (2) lipase-digested sesame oil; (3) undigested sesame oil; (4) five additional common vegetable oils; (5) mineral oil. Linoleic inhibited the in vitro growth of all three malignant human adenocarcinoma cell lines. The normal cell line showed dramatically less inhibition of growth. Lipase-digested sesame oil (LDSO) and undigested sesame oil (UDSO) produced greater inhibition of growth of all three malignant cell lines than of the normal cells. Five other common vegetable oils containing various amounts of PUFAs such as corn, soybean, safflower, olive and coconut oils, all in their lipase-digested form, were found to dramatically inhibit the growth of the HT-29 malignant human cell line. Undigested olive and safflower oils also inhibited the HT-29 cells although not as markedly as the lipase-digested oils. Mineral oil did not inhibit the growth of HT-29 cells. Both lauric and , which are saturated fatty acids found in abundance in coconut oil inhibits the HT-29 cells more strongly than linoleic , while oleic did not inhibit. We conclude that many vegetable oils including sesame contain in vitro antineoplastic properties and that this finding warrants further investigation both in vitro and in vivo to assess their possible chemotherapeutic potential.

Keyword: colon cancer

Role of fatty acids in malignancy and visual impairment: epidemiological evidence and experimental studies.

International variation in breast and incidence is positively related to total fat intake. However, total fat consists of different fatty families, e.g., saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and n-3 and n-6 polyunsaturated fatty acids (PUFAs). Epidemiological evidence and experimental studies suggest that these fatty families have different effects on breast and carcinogenesis. Therefore the action of each fatty on carcinogenesis should be evaluated separately. Although it is difficult to establish firm conclusions on the effect of each fatty in human epidemiological studies, experimental studies on animals and cultured cells suggest that n-6 PUFAs (linoleic and arachidonic ) may have a tumor promoting effect, while n-3 PUFAs (eicosapentaenoic , docosahexaenoic and alpha-linolenic ) and conjugated fatty acids (CFAs; a mixture of positional and geometric isomers of PUFAs with conjugated double bonds) exert an inhibitory effect on tumor growth. SFAs such as and stearic show little or no tumor promoting effect, and the action of oleic , a MUFA, is inconclusive. In addition to regulation of abnormal cell growth seen in cancers, fatty acids also control cell loss seen in degenerative eye diseases, such as degeneration of lens material in cataract and degeneration of photoreceptor cells in retinitis pigmentosa. Experiments suggest that n-6 PUFAs cause deleterious effects, while n-3 PUFAs result in beneficial effects on the lens and retina. In particular, docosahexaenoic is known to be effective in rescuing photoreceptor cells from damage. Thus, understanding the function of each fatty is likely to be important for making progress in treating these and other diseases.

Keyword: colon cancer

Type 2 Diabetes Promotes Cell Centrosome Amplification via AKT-ROS-Dependent Signalling of ROCK1 and 14-3-3σ.

Type 2 diabetes is associated with oxidative stress and DNA damage which can cause centrosome amplification. Thus, the study investigated centrosome amplification in type 2 diabetes and the underlying mechanisms.Centrosome numbers in human peripheral blood mononuclear blood cells (PBMC) from healthy subjects and patients with type 2 diabetes were compared to access the association between type 2 diabetes and centrosome amplification. cells were used to investigate the molecular mechanisms underlying the centrosome amplification triggered by high glucose, insulin and . Western blot analysis was used to quantify the level of protein and protein phosphorylation. Immunofluorescent staining was performed to detect centrosomes. ROS was quantified using flow cytometry technique. Transcriptpmic profiling was performed using Illumina HiSeqTM500 platform.We found that centrosome amplification was increased PBMC from the type 2 diabetic patients, which correlated with the levels of fasting blood glucose and HbA1c. High glucose, insulin and , alone or in combinations, induced ROS production and centrosome amplification. Together, they increased AKT activation as well as the expression, binding and centrosome translation of ROCK1 and 14-3-3σ. Results from further analyses showed that AKT-ROS-dependent upregulations of expression, binding and centrosome translocation of ROCK1 and 14-3-3σ was the molecular pathway underlying the centrosome amplification in vitro triggered by high glucose, insulin and . Moreover, the key in vitro molecular signalling events activated by high glucose, insulin and were verified in PBMC from the patients with type 2 diabetes.Our results show that type 2 diabetes promotes cell centrosome amplification, and suggest that the diabetic pathophysiological factors-activated AKT-ROS-dependent signalling of ROCK1 and 14-3-3σ is the underlying molecular mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: colon cancer

Isolation of fatty acids with anticancer activity from Protaetia brevitarsis larva.

In this study, biologically active compounds were isolated from Protaetia brevitarsis larva (PBL) by dichloromethane extraction. The dichloromethane extract from PBL was highly cytotoxic to various cells. From a silica gel column chromatograpy of this extract, we obtained four fractions (F-2, F-4, F-5 and F-7) having apoptosis-inducing activity. These fractions induced DNA ladder and caspase-3 activation during apoptosis in 26 tumor cells. In 1H and 13C NMR and mass spectral analysis of the fraction F-2 showing the highest apoptosis-inducing activity, we found that the fraction was composed of three free fatty acids such as , (Z)-9-octadecenoic and octadecenoic . These results indicate that the dichloromethane extract of PBL includes anticancer components composed of at least three fatty acids, and apoptosis-inducing activity of the extract was mediated by caspase-3 activation in tumor cells.

Keyword: colon cancer

Effects of fatty acids on the growth of Caco-2 cells.

Epidemiological studies suggest that polyunsaturated fatty acids may protect against colorectal neoplasia. In order to explore this observation, cell proliferation and viability, lipid composition, membrane fluidity, and lipid peroxidation were measured in Caco-2 cells after 48h incubation with various fatty acids. Saturated and monounsaturated fatty acids incorporated less well in the membranes than polyunsaturated fatty acids (PUFAs). All of the PUFAs tested had an inhibitory effect on cell proliferation/viability whereas the saturated and monounsaturated fatty acids did not. Addition of had no significant effect on membrane fluidity whereas unsaturated fatty acids increased membrane fluidity in a dose-dependent manner. PUFAs strongly increased tumor cell lipid peroxidation in a dose-dependent manner. Saturated and monounsaturated fatty acids increased lipid peroxidation in this cell line only at high concentration. Preincubation of Caco-2 cells with vitamin E prevented the inhibition of proliferation/viability, the elevation of the MDA concentration and the increased membrane fluidity induced by PUFAs. Our data indicate that PUFAs are potent inhibitors of the growth of cells in vitro.

Keyword: colon cancer

Mitochondrial hexokinase from differentiated and undifferentiated HT29 cells: effect of some metabolites on the bound/soluble equilibrium.

1. Solubilization of mitochondrial bound hexokinase (HK), which represents 75-80% of the total enzyme activity in the cells, was investigated in freshly isolated mitochondria from undifferentiated (Glc+) or differentiated (Glc-) HT29 adenocarcinoma cells. In both models, the bound HK is almost completely released in vitro by 100 microM glucose 6-P (G 6-P). 2. Free ATP (5 mM) or palmitate (800 microM) produce a partial solubilization of bound HK, more markedly in the case of Glc- mitochondria. 3. Glucose or glucose 1-P are found unable to solubilize bound HK. Glucose 1,6-P2, 2-deoxyglucose 6-P or glucosamine 6-P can solubilize the enzyme but are less efficient than G 6-P. 4. Mg2+ and Pi are found to counteract the glucose 6-P induced solubilization of HK in both types of mitochondria. Taking into account the intracellular concentrations of these ions, this could in part explain why, in HT29 cells, HK is predominantly bound to the mitochondria.

Keyword: colon cancer

N-(4-Hydroxyphenyl)retinamide increases dihydroceramide and synergizes with dimethylsphingosine to enhance cell killing.

Fenretinide [N-(4-hydroxyphenyl)retinamide (4-HPR)] is cytotoxic in many cell types. Studies have shown that elevation of ceramide species plays a role in 4-HPR cytotoxicity. To determine 4-HPR activity in a multidrug-resistant cell line as well as to study ceramide metabolism, MCF-7/AdrR cells (redesignated NCI/ADR-RES) were treated with 4-HPR and sphingolipids were analyzed. TLC analysis of cells radiolabeled with [3H] showed that 4-HPR elicited a dose-responsive increase in radioactivity migrating in the ceramide region of the chromatogram and a decrease in cell viability. Results from liquid chromatography/electrospray tandem mass spectrometry revealed large elevations in dihydroceramides (N-acylsphinganines), but not desaturated ceramides, and large increases in complex dihydrosphingolipids (dihydrosphingomyelins, monohexosyldihydroceramides), sphinganine, and sphinganine 1-phosphate. To test the hypothesis that elevation of sphinganine participates in the cytotoxicity of 4-HPR, cells were treated with the sphingosine kinase inhibitor d-erythro-N,N-dimethylsphingosine (DMS), with and without 4-HPR. After 24 h, the 4-HPR/DMS combination caused a 9-fold increase in sphinganine that was sustained through +48 hours, decreased sphinganine 1-phosphate, and increased cytotoxicity. Increased dihydrosphingolipids and sphinganine were also found in HL-60 leukemia cells and HT-29 cells treated with 4-HPR. The 4-HPR/DMS combination elicited increased apoptosis in all three cell lines. We propose that a mechanism of 4-HPR-induced cytotoxicity involves increases in dihydrosphingolipids, and that the synergy between 4-HPR and DMS is associated with large increases in cellular sphinganine. These studies suggest that enhanced clinical efficacy of 4-HPR may be realized through regimens containing agents that modulate sphingoid base metabolism.

Keyword: colon cancer

Alcohol Consumption-Related Metabolites in Relation to Colorectal and Adenoma: Two Case-Control Studies Using Serum Biomarkers.

Alcohol is a known carcinogen that may be associated with colorectal . However, most epidemiologic studies assess alcoholic beverage consumption using self-reported data, leading to potential exposure misclassification. Biomarkers of alcohol consumption may provide an alternative, complementary approach that reduces misclassification and incorporates individual differences in alcohol metabolism. Therefore, we evaluated the relationship between previously identified alcohol consumption-related metabolites and colorectal and adenoma using serum metabolomics data from two studies. Data on colorectal were obtained from a nested case-control study of 502 US adults (252 cases, 250 controls) within the Prostate, Lung, Colorectal, and Ovarian Screening Trial. Data on colorectal adenoma were obtained from a case-control study of 197 US adults (120 cases, 77 controls) from the Navy Adenoma Study. Unconditional multivariable logistic regression models were fit to calculate odds ratios (OR) and 95% confidence intervals (CI) for eight alcohol consumption-related metabolites identified in a previous analysis: ethyl glucuronide; 4-androstene-3beta,17beta-diol disulfate 1; 5-alpha-androstan-3beta,17beta-diol disulfate; 16-hydroxypalmitate; bilirubin (E,Z or Z,E); cyclo (-leu-pro); dihomo-linoleate (20:2n6); and palmitoleate (16:1n7). We found no clear association between these alcohol consumption-related metabolites and either endpoint. However, we did observe an inverse association between cyclo (-leu-pro) and colorectal adenoma that was only observed in the highest metabolite quantile (OR 4th vs. 1st Quantile = 0.30, 95% CI: 0.12-0.78; P-trend = 0.047), but no association for colorectal . In conclusion, there were no adverse associations between alcohol consumption-related metabolites and colorectal or adenoma.

Keyword: colon cancer

Role of acylethanolamides in the gastrointestinal tract with special reference to food intake and energy balance.

Acylethanolamides (AEs) are a group of lipids occurring in both plants and animals. The best-studied AEs are the endocannabinoid anandamide (AEA), the anti-inflammatory compound palmitoylethanolamide (PEA), and the potent anorexigenic molecule oleoylethanolamide (OEA). AEs are biosynthesized in the gastrointestinal tract, and their levels may change in response to noxious stimuli, food deprivation or diet-induced obesity. The biological actions of AEs within the gut are not limited to the modulation of food intake and energy balance. For example, AEs exert potential beneficial effects in the regulation of intestinal motility, secretion, inflammation and cellular proliferation. Molecular targets of AEs, which have been identified in the gastrointestinal tract, include cannabinoid CB(1) and CB(2) receptors (activated by AEA), transient receptor potential vanilloid type 1 (TRPV1, activated by AEA and OEA), the nuclear receptor peroxisome proliferators-activated receptor-alpha (PPAR-alpha, activated by OEA and, to a less extent, by PEA), and the orphan G-coupled receptors GPR119 (activated by OEA) and GPR55 (activated by PEA and, with lower potency, by AEA and OEA). Modulation of AE levels in the gut may provide new pharmacological strategies not only for the treatment of feeding disorders but also for the prevention or cure of widespread intestinal diseases such as inflammatory bowel disease and .

Keyword: colon cancer

Injury induced by fatty acids or bile in isolated human colonocytes prevented by calcium.

Measurement of the modulation of the growth fraction of isolated normal colonocytes from adult subjects in primary monolayer culture was used as a sensitive quantitative assay to evaluate toxic effects of several endogenous compounds found within the . This assay was used to study the role of CaCl2 in blocking cell injury. When added simultaneously with the injurious agent, 5-10 mM CaCl2 blocked the toxicity of physiological concentrations of deoxycholic , oleic , and linoleic .

Keyword: colon cancer

n-6 and n-3 polyunsaturated fatty acids differentially modulate oncogenic Ras activation in colonocytes.

Ras proteins are critical regulators of cell function, including growth, differentiation, and apoptosis, with membrane localization of the protein being a prerequisite for malignant transformation. We have recently demonstrated that feeding fish oil, compared with corn oil, decreases Ras membrane localization and reduces tumor formation in rats injected with a carcinogen. Because the biological activity of Ras is regulated by posttranslational lipid attachment and its interaction with stimulatory lipids, we investigated whether docosahexaenoic (DHA), found in fish oil, compared with linoleic (LA), found in corn oil, alters Ras posttranslational processing, activation, and effector protein function in young adult mouse cells overexpressing H-ras (YAMC-ras). We show here that the major n-3 polyunsaturated fatty (PUFA) constituent of fish oil, DHA, compared with LA (an n-6 PUFA), reduces Ras localization to the plasma membrane without affecting posttranslational lipidation and lowers GTP binding and downstream p42/44(ERK)-dependent signaling. In view of the central role of oncogenic Ras in the development of , the finding that n-3 and n-6 PUFA differentially modulate Ras activation may partly explain why dietary fish oil protects against development.

Keyword: colon cancer

Dietary soy sphingolipids suppress tumorigenesis and gene expression in 1,2-dimethylhydrazine-treated CF1 mice and ApcMin/+ mice.

Dietary supplementation with milk sphingolipids inhibits tumorigenesis in CF1 mice treated with a carcinogen [1,2-dimethylhydrazine (DMH)] and in multiple intestinal neoplasia (Min) mice, which develop intestinal tumors spontaneously. Plant sphingolipids differ structurally from those of mammals [soy glucosylceramide (GlcCer) consists predominantly of a 4,8-sphingadiene backbone and alpha-hydroxy-], which might affect their bioactivity. Soy GlcCer was added to the AIN-76A diet (which contains <0.005% sphingolipid) to investigate whether it would also suppress tumorigenesis in these mouse models. Soy GlcCer reduced cell proliferation in the upper half of the crypts in mice treated with DMH by 50 and 56% (P < 0.05) at 0.025 and 0.1% of the diet (wt/wt), respectively, and reduced the number of aberrant crypt foci (an early marker of carcinogenesis) by 38 and 52% (P < 0.05). Min mice fed diets containing 0.025 and 0.1% (wt/wt) soy GlcCer developed 22 and 37% fewer adenomas (P < 0.05), respectively. The effects of dietary sphingolipids on gene expression in the intestinal mucosal cells of Min mice were analyzed using Affymetrix GeneChip microarrays. Soy GlcCer affected the expression of 96 genes by > or = 2-fold in a dose-dependent manner, increasing 32 and decreasing 64. Decreases in the mRNA expression of two transcription factors associated with , hypoxia-induced factor 1 alpha (HIF1 alpha) and transcription factor 4 (TCF4), were confirmed by quantitative RT-PCR. In conclusion, soy GlcCer suppressed tumorigenesis in two mouse models; hence, plant sphingolipids warrant further investigation as inhibitors of . Because soy contains relatively high amounts of GlcCer, sphingolipids may partially account for the anticancer benefits attributed to soy-based foods.

Keyword: colon cancer

Tissue-Specific Distribution of Sphingomyelin Species in Pork Chop Revealed by Matrix-Assisted Laser Desorption/Ionization-Imaging Mass Spectrometry.

Sphingomyelin (SM) species are major sphingolipids in pork meat that affect quality parameters, such as health benefits due to their protective properties against chronic diseases; however, their spatial distribution remains unclear. We used matrix-assisted laser desorption/ionization (MALDI)-imaging mass spectrometry (IMS) to investigate the distribution and composition of SM species in pork chop consisting of longissimus thoracis et lumborum muscle (loin), intermuscular fat tissue, transparent tissue, and spinalis muscle. Four SM species were identified by liquid chromatography-electrospray ionization-tandem MS (MS/MS) and MALDI-MS/MS and visualized using MALDI-IMS. SM species containing stearic were predominantly distributed in the loin and spinalis muscle, whereas SM species containing , lignoceric, and nervonic acids were predominantly distributed in transparent tissue. These results indicated that the distribution of SM species differed among the pork tissues, depending on the tissue-specific fatty composition. The total amount including all identified SM species was higher in the loin than in spinalis muscle. Pork is reportedly associated with increased risk for chronic diseases due to the high amount of heme iron. From the observation of color, the amount of heme iron was lower in loin than in spinalis muscle. Thus, the degree of risk for chronic diseases might be lower in the loin than in spinalis muscle. This is the first report on the tissue-specific distribution of SM species in meat at a microscopic resolution using IMS. MALDI-IMS analysis may be useful in assessing the association between SM species and quality parameters of pork meat. PRACTICAL APPLICATION: Sphingomyelin (SM) species are major sphingolipids in pork meat. SM species affect quality parameters such as health benefits due to their protective properties against and atherosclerosis. Matrix-assisted laser desorption/ionization-imaging mass spectrometry analysis combined with liquid chromatography-electrospray ionization-tandem mass spectrometry is a suitable method to directly investigate the distribution and composition of SM species at microscopic level among different tissues of pork meat. Therefore, this method is useful to assess the SM species-induced health effect of different tissues of pork meat.© 2019 Institute of Food Technologists®.

Keyword: colon cancer

A bitter melon extract inhibits the P-glycoprotein activity in intestinal Caco-2 cells: monoglyceride as an active compound.

P-glycoprotein (P-gp) is a 170 kDa membrane protein that belongs to the ATP-binding cassette (ABC) transporter superfamily. In normal tissues, P-gp functions as an ATP-dependent efflux pump that excretes highly hydrophobic xenobiotic compounds, playing an important role in protecting the cells/tissues from xenobiotics. In the present study, chemical substances that could directly modulate the intestinal P-gp activity were searched in vegetables and fruits. By using human intestinal epithelial Caco-2 cells as a model of the small intestinal cells, we observed that a bitter melon fraction extracted from 40% methanol showed the greatest increase of the rhodamine-123 accumulation by Caco-2 cells. Inhibitory compounds in the bitter melon fraction were then isolated by HPLC using Pegasil C4 and Pegasil ODS columns. The HPLC fraction having the highest activity was analyzed by (1)H-NMR and FAB-MS, and the active compound was identified as 1-monopalmitin. It is interesting that certain types of monoglyceride might be involved in the drug bioavailability by specifically inhibiting the efflux mediated by P-gp.

Keyword: colon cancer

Acetyl-CoA carboxylase-alpha inhibitor TOFA induces human cell apoptosis.

Acetyl-CoA carboxylase-alpha (ACCA) is a rate-limiting enzyme in long chain fatty synthesis, playing a critical role in cellular energy storage and lipid synthesis. ACCA is upregulated in multiple types of human cancers and small interfering RNA-mediated ACCA silencing in human breast and prostate cells results in oxidative stress and apoptosis. This study reports for the first time that TOFA (5-tetradecyloxy-2-furoic ), an allosteric inhibitor of ACCA, is cytotoxic to lung cells NCI-H460 and carcinoma cells HCT-8 and HCT-15, with an IC(50) at approximately 5.0, 5.0, and 4.5 microg/ml, respectively. TOFA at 1.0-20.0 microg/ml effectively blocked fatty synthesis and induced cell death in a dose-dependent manner. The cell death was characterized with PARP cleavage, DNA fragmentation, and annexin-V staining, all of which are the features of the apoptosis. Supplementing simultaneously the cells with acids (100 microM), the end-products of the fatty synthesis pathway, prevented the apoptosis induced by TOFA. Taken together, these data suggest that TOFA is a potent cytotoxic agent to lung and cells, inducing apoptosis through disturbing their fatty synthesis.

Keyword: colon cancer

Identification of Palmitoylated Transitional Endoplasmic Reticulum ATPase by Proteomic Technique and Pan Antipalmitoylation Antibody.

Protein palmitoylation plays a significant role in a wide range of biological processes such as cell signal transduction, metabolism, apoptosis, and carcinogenesis. For high-throughput analysis of protein palmitoylation, approaches based on the acyl-biotin exchange or metabolic labeling of azide/alkynyl-palmitate analogs are commonly used. No palmitoylation antibody has been reported. Here, the palmitoylated proteome of human cell lines SW480 was analyzed via a TS-6B-based method. In total, 151 putative palmitoylated sites on 92 proteins, including 100 novel sites, were identified. Except for 3 known palmitoylated transmembrane proteins, ATP1A1, ZDHHC5, and PLP2, some important proteins including kinases, ion channels, receptors, and cytoskeletal proteins were also identified, such as CLIC1, PGK1, PPIA, FKBP4, exportin-2, etc. More importantly, the pan antipalmitoylation antibody was developed and verified for the first time. Our homemade pan antipalmitoylation antiserum could differentiate well protein palmitoylation from mouse brain membrane fraction and SW480 cells, which affords a new technique for analyzing protein palmitoylation by detecting the moiety directly. Furthermore, the candidate protein transitional endoplasmic reticulum ATPase (VCP) identified in SW480 cells was validated to be palmitoylated by Western blotting with anti-VCP antibody and the homemade pan antipalmitoylation antibody.

Keyword: colon cancer

Effects of specific fatty acids on cell transformation induced by an activated c-H-ras oncogene.

An increase in dietary lipid has been associated with an increase in the development of certain forms of , notably breast and , both in experimental animal studies and in human epidemiology studies. The underlying mechanisms are not, however, known with certainty. In the present studies we have examined whether certain specific fatty acids (FA) might act by enhancing the role of an activated oncogene in a model cell culture system. We found that when the rat fibroblast cell line Rat 6 was transfected with an activated human c-H-ras oncogene and the cells subsequently grown in medium supplemented with myristic , or stearic (20-80 microM) there was a marked enhancement of the number of transformed foci obtained. On the other hand arachidonic had a marked inhibitory effect in this transformation assay. However, this inhibitory effect can be partially reversed by indomethacin, an inhibitor of cyclo-oxygenase, at dose response manner. Control studies indicated that these results were not simply due to the effects of the FAs on growth of the Rat 6 cells or the process of transfection per se. Lipid analyses of cells grown in the presence of stearic indicated that the added FA was extensively incorporated into the major lipid classes of the cell and produced transient changes in lipid composition. This simple cell culture system may be useful for elucidating the mechanisms by which various dietary lipids and nutritional factors influence the carcinogenic process.

Keyword: colon cancer

Effects of fatty acids on liver metastasis of ACL-15 rat cells.

The effects of eicosapentaenoic [EPA; n-3 polyunsaturated fatty (PUFA)], linoleic (LA; n-6 PUFA), and (PA; saturated fatty ) on 1,2-dimethylhydrazine-induced F344 rat carcinoma cells (ACL-15) were investigated in vivo and in vitro. The number and size of liver metastatic foci via a superior mesenteric vein injection of ACL-15 cells in F344 rats were significantly inhibited in the EPA-treated group compared with the LA-treated group (p < 0.01); the PA-treated animals and those fed commercial rodent chow (standard diet) demonstrated intermediate values. In a dot immunoblotting assay, vascular cell adhesion molecule 1 expression on ACL-15 cells was downregulated by EPA-ethyl ester treatment and upregulated by LA-ethyl ester treatment compared with the untreated control cells, whereas the expression of matrix metalloproteinase 1 and 2 was not influenced by the fatty ethyl esters. In a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, EPA-ethyl ester suppressed ACL-15 cell growth in a schedule-dependent manner, and LA-ethyl ester showed schedule-dependent stimulation. In contrast, PA demonstrated no regulatory effect on cell growth at lower concentrations (< or = 5 mg/ml) but concentration-dependent inhibition at higher concentrations. According to our in vivo cell kinetic study, the difference in tumor growth at the metastatic site was due to different tumor cell proliferation rates; the cell loss rate was not altered. Therefore, the inhibitory effect of liver metastasis on ACL-15 cells by EPA can be explained by a decreased ability of tumor cell adhesion to the capillary bed (low expression of vascular cell adhesion molecule 1) and a lower potential of tumor cell proliferation (low mitotic rate) at the secondary site.

Keyword: colon cancer

Further insights into chemical characterization through GC-MS and evaluation for anticancer potential of Dracaena draco leaf and fruit extracts.

The present study reports for the first time the amino and fatty compositions and the antitumoral activity of aqueous extracts obtained from Dracaena draco L. leaf and fruit. Metabolite profiles were determined by gas chromatography-ion trap-mass spectrometry (GC-IT-MS), with several amino acids, , linolenic and stearic being identified in the leaf extract, and only proline, oleic and stearic in the fruit extract. The in vitro antiproliferative activities of the extracts were tested against human (Caco-2), kidney (A-498), and liver (HepG2) cell lines. In addition, primary cultures of normal and cancerous renal cells derived from kidney patients were treated with D. draco extracts (0-400 μg/mL). Antiproliferative and cytotoxic effects were determined by the MTT assay. D. draco extracts inhibited proliferation of human and renal tumor cells in vitro, whereas no or weak effect was observed in HepG2 cells. Compared to the fruit extract, D. draco leaf extract exhibited stronger antiproliferative activity against all cells. Our results indicate that D. draco, particularly the leaf, may be useful as a chemopreventive and/or chemotherapeutic agent for and kidney cancers.Copyright © 2012 Elsevier Ltd. All rights reserved.

Keyword: colon cancer

Fatty acylation of heparan sulfate proteoglycan from human carcinoma cells.

A number of transmembrane proteins have been recently reported to be modified by the covalent addition of saturated fatty acids which may contribute to membrane targeting and specific protein-lipid interactions. Such modifications have not been reported in cell-associated heparan sulfate proteoglycans, although these macromolecules are known to be hydrophobic. Here, we report that a cell surface heparan sulfate proteoglycan is acylated with both myristate and palmitate, two long-chain saturated fatty acids. When carcinoma cells were labeled with [3H]myristic , a significant proportion of the label was shown to be specifically incorporated into the protein core of the proteoglycan. Characterization of fatty acyl moiety in the purified proteoglycan by reverse-phase high pressure liquid chromatography revealed that approximately 60% of the covalently bound fatty acids was myristate. We further show that this relatively rare 14-carbon fatty was bound to the protein core via a hydroxylamine- and alkali-resistant amide bond. The remaining 40% was the more common 16-carbon palmitate, which was bound via a hydroxylamine- and alkali-sensitive thioester bond. Palmitate appeared to be added post-translationally and derived in part from intracellular elongation of myristate, a process that occurred within the first two hours and was insensitive to inhibition of protein synthesis. Acylation of heparan sulfate proteoglycan represents a novel modification of this gene product and could play a role in a number of biological functions including specific interactions with membrane receptors and ligand stabilization.

Keyword: colon cancer

Increased carnitine-dependent fatty uptake into mitochondria of human cells induces apoptosis.

Carnitine-dependent fatty import into mitochondria and beta-oxidation seem to be impaired in tumor cells. In the present study we show that a supply of palmitoylcarnitine together with L-carnitine potently induces apoptosis in HT-29 human cells as a consequence of accelerated fatty oxidation. Caspase-3-like activities, measured by the cleavage rate of a fluorogenic tetrapeptide substrate and nuclear fragmentation determined after DNA labeling in fixed cells by fluorescence microscopy, served as indicators of apoptosis. Neither L-carnitine nor palmitoylcarnitine alone were able to increase caspase-3-like activities and DNA fragmentation, but when provided together, apoptosis occurred. That exogenous carnitine was indeed able to enhance fatty uptake into mitochondria was demonstrated by an increased influx of a fluorescent analog. Enhanced fatty availability in mitochondria led to an increased generation of O*2-, as detected by a O*2- -sensitive fluorogenic dye, indicating oxidation of delivered substrates. Benzoquinone, an O*2- scavenger, blocked O*2- generation and prevented apoptosis as initiated by the combination of palmitoylcarnitine and carnitine. The lack of effect of the ceramide synthesis inhibitor fumonisin on palmitoylcarnitine/carnitine-induced apoptosis further supports the notion that apoptotic cell death is specifically due to fatty oxidation. In contrast to HT-29 cells, nontransformed human colonocytes did not respond to exogenous palmitoylcarnitine/carnitine and no apoptosis was observed. In conclusion, our studies provide evidence that a limited mitochondrial fatty import in human cells prevents high rates of mitochondrial O*2- production and protects cells from apoptosis that can be overcome by an exogenous carnitine supply.

Keyword: colon cancer

Hexadecanoic and neuraminic incorporations in two rat carcinoma cell lipids: selective influence of 1-O-octadecyl 2-O-methyl-3-phosphocholine on glycerolipid and ganglioside biosynthesis.

[3H] hexadecanoic and N-acetyl [14C] neuraminic acids were incorporated in glycerolipids or gangliosides of 2 rat carcinoma cell lines, having (PRO cells), or not (REG cells) invasive capacities when inoculated in syngeneic BD IX rats. The cells were cultured (48 h) in presence of 1-0-octadecyl-2-0-methyl-3-phosphocholine (ET 18-0-CH3) 20 or 40 microM, which, on transformed cells, inhibits the cell growth, modifies the glycerolipid biosynthesis, and activates the sialyltransferases. ET 18-0-CH3 20 microM activated, in PRO and in REG cells the incorporation of [3H] hexadecanoate in monosialogangliosides (1.45 fold compared to controls), but not in disialogangliosides and the distribution of this fatty between monosialo- (82%) and disialogangliosides (18%) was unchanged with controls. After [14C] neuraminic labelings, and for control experiments, the total radioactivities in gangliosides, in PRO cells, were twice higher than in REG cells, a difference which, probably, reflects the ganglioside content. ET 18-0-CH3 20 microM did not increase the incorporation of the [14C] neuraminic in PRO and in REG cells, and did not change its distribution between monosialo (70-80% for controls and experiments with ET 18-0-CH3) and disialogangliosides (20-30%). Similar results were obtained with ET 18-0-CH3 40 microM for the distribution of [14C] neuraminic in monosialo- and disialogangliosides. Whatever the precursor, the trisialogangliosides were never radiolabeled. Analysis of the [3H] glycerolipids (the main radiolabeled lipid classes in controls were: phosphatidylcholines, triglycerides, sphingomyelins and phosphatidyl-inositols) revealed that ET 18-0-CH3, compared to controls, did not activate the incorporation of [3H] hexadecanoate in total glycerolipids (PRO or REG cells). It activated (3 fold) its incorporation in triglyerides, inhibited it (0.5-0.6 fold) in phosphatidylcholines, sphingomyelins and phosphatidyl-inoditols and all these most noticeable differences were observed in PRO and in REG cells. These findings reflect the impossibility of ET 18-0-CH3 to activate the sialyltransferases during the ganglioside biosynthesis in carcinoma cells, while it modified ceramide, glycerophospholipid and neutral glycerolipid biosynthesis.

Keyword: colon cancer

PPARγ promotes diabetes-associated centrosome amplification via increasing the expression of SKA1 directly at the transcriptional level.

We have\xa0recently published that type 2 diabetes can induce cell centrosome amplification due to the action of high glucose, , and insulin, and ROCK1 and 14-3-3σ are signal mediators. In this study, we further investigated the molecular mechanisms of the centrosome amplification in HCT116 cells. Treatment of the cells with high glucose, , and insulin increased the expression of peroxisome proliferator-activated receptor γ (PPARγ) as well as the spindle and kinetochore associated protein 1 (SKA1), knockdown of each of which resulted in the inhibition of the treatment-triggered centrosome amplification. Knockdown of PPARγ inhibited the treatment-evoked increase in the SKA1 level, whereas knockdown of SKA1 did not modify the treatment-increased PPARγ level. We found a predicted binding site for PPARγ in the promoter region of the SKA1 gene from the JASPAR database. Experimental results showed that the treatment increased the messenger RNA level of SKA1, which could be inhibited by PPARγ chemical inhibitor or small interfering RNA. Moreover, we were able to show that PPARγ could bind to the binding site in the SKA1 gene promoter, which was increased by the experimental treatment. In conclusion, it is suggested that the pathophysiological factors in type 2 diabetes, high glucose, , and insulin, induce the cell centrosome amplification through the PPARγ-SKA1 pathway, in which PPARγ increases the expression of SKA1 via directly enhancing the SKA1 gene transcription.© 2019 Wiley Periodicals, Inc.

Keyword: colon cancer

Neutral glycolipid composition in human .

Neutral glycolipid composition of human gastric and was studied in comparison to healthy mucosa. The following compounds were identified: galactosylceramide, glucosylceramide, trihexosylceramide, globoside, small amounts of neotetraosylceramide and fucoglycolipids. The latter comprised both 3-0- and 4-0-substituted residues of N-acetylglucosamine. In tissue the contents of lactosylcermide and of fucoglycolipids were increased. Fatty compositions of neutral glycolipids derived from both and control samples were similar. The most abundant fatty acids were and nervonic acids.

Keyword: colon cancer

Dietary and biomarker estimates of fatty acids and risk of colorectal .

The associations between intake of or circulating fatty acids and risk of colorectal (CRC) are unclear. We examined prospectively the associations between dietary or biomarker fatty acids and CRC. For 41,514 men and women, aged 40-69 years, baseline (1990-94) dietary intakes of fatty acids were estimated using a food frequency questionnaire and plasma phospholipid (PPL) fatty acids were measured for 4,205 participants including 395 CRC cases, according to a case-cohort design. Hazard ratios were computed using Cox regression adjusting for education, alcohol intake, smoking status, physical activity and total energy intake; and stratified for gender, ethnicity and family history of , with age as the time scale. We assessed the heterogeneity of associations with and rectal cancers. PPL saturated fatty acids (SFAs) were positively associated with CRC risk, while total n-3 polyunsaturated fatty acids (PUFA) and long chain marine n-3 PUFAs showed inverse associations, significant only for 22:5 n-3. No significant associations were observed for dietary fatty intakes but positive associations with CRC of borderline significance were seen for both dietary and PPL linoleic . Positive associations with dietary (16:0), MUFAs and n-6 PUFAs were seen for rectal but not cancers. PPL 22:6 n-3 was inversely associated with rectal . Limiting intakes of SFAs and MUFAs could be assisted by following existing guidelines to limit red and processed meats which are important sources in the Australian diet. Our observations regarding linoleic should be examined further.© 2015 UICC.

Keyword: colon cancer

Effect of dietary tetradecylthioacetic on growth studied by dynamic contrast enhanced MRI.

To evaluate the effects of diets supplemented with the modified fatty tetradecylthioacetic (TTA) and fish protein hydrolysate (FPH) on tumor growth of the human cell line SW620, and to investigate the properties of tumor vasculature by dynamic contrast-enhanced MRI in a human tumor xenograft.SW620 cells were grown in vitro in presence of TTA and and proliferation was measured by thymidine incorporation. The xenograft study in mice was performed with four distinct diets: (a) control diet; (b) diet with TTA; (c) diet with TTA and FPH; and (d) diet with FPH. SW620 cells were injected subcutaneously, and dynamic contrast enhanced (DCE) MRI was performed on a Bruker BioSpec 7T system. The data was analyzed by two-compartment modeling of the contrast enhancement, initial area under the curve (IAUC) and by use of relative signal intensity (RSI) distributions.The in vitro cell studies revealed that TTA reduced tumor cell proliferation as a function of both dose and time. The in vivo tumor growth was significantly reduced for the two groups fed TTA, as compared to the control group. The mean 10(th) percentile RSI, v(e) and IAUC for the TTA group were significant higher than for the control group.This study confirms the growth inhibitory effects of TTA, both in vitro and in vivo, in a model. The analysis of DCE-MRI data showed that TTA influences the vascular properties of the tumor in addition to the growth.

Keyword: colon cancer

Fatty esterification during differentiation of the human intestinal cell line Caco-2.

The Caco-2 human intestinal cell line was used to examine fatty esterification during development of the enterocytic phenotype. Acyl-CoA synthetase activity increased approximately 40%, and the incorporation of into triacylglycerol relative to phosphatidylcholine increased nearly 2-fold during Caco-2 differentiation. A rate-limiting enzyme activity in the glycerol 3-phosphate pathway of triacylglycerol synthesis, glycerol-3-phosphate acyltransferase, was at levels comparable with rat jejunum and remained unchanged during differentiation. In contrast, the activity of monoacylglycerol acyltransferase, which is unique to the monoacylglycerol pathway of triacylglycerol synthesis, was present at < 7% of the levels in rat jejunum. Further analysis of the glycerol 3-phosphate pathway showed that the rate-limiting enzyme activities for diacylglycerol conversion to triacylglycerol, diacylglycerol acyltransferase, and phosphatidylcholine, CTP:phosphocholine cytidylyltransferase, increased 2-3-fold and decreased approximately 40%, respectively, during Caco-2 differentiation. In addition, a 2-fold increase in cellular diacylglycerol mass was observed during enterocytic conversion. These data indicate that fatty esterification to triacylglycerol in Caco-2 cells occurs primarily via the glycerol 3-phosphate pathway. Furthermore, the differentiation-dependent increase in fatty esterification to triacylglycerol relative to phosphatidylcholine appears to result from increased utilization of diacylglycerol to synthesize triacylglycerol and a concomitant decrease in diacylglycerol utilization for phosphatidylcholine synthesis.

Keyword: colon cancer

PCNA and JNK1-Stat3 pathways respectively promotes and inhibits diabetes-associated centrosome amplification by targeting at the ROCK1/14-3-3σ complex in human HCT116 cells.

We have recently reported that type 2 diabetes promotes centrosome amplification via enhancing the expression, biding, and centrosome translocation of rho-associated coiled-coil containing protein kinase 1 (ROCK1)/14-3-3σ complex in HCT116 cells. In the functional proteomic study, we further investigated the molecular pathways underlying the centrosome amplification using HCT116 cells. We found that treatment of HCT116 cells with high glucose, insulin, and triggered the centrosome amplification and increased the expressions of proliferating cell nuclear antigen (PCNA), nucleophosmin (NPM), and 14-3-3σ. Individual knockdown of PCNA, NPM, or 14-3-3σ inhibited the centrosome amplification. Knockdown of PCNA inhibited the treatment-increased expression of ROCK1, whereas knockdown of ROCK1 did not affect the PCNA expression. High glucose, insulin, and also increased the expressions of c-Jun N-terminal kinase-1 (JNK1) and signal transducer and activator of transcription 3 (Stat3), individual knockdown of which upregulated the treatment-increased expression of 14-3-3σ and promoted the centrosome amplification. In contrast, overexpression of JNK1 inhibited the centrosome amplification. Knockdown of Stat3 enhanced the centrosome translocation of 14-3-3σ. Moreover, we showed that knockdown of JNK1 inhibited the treatment-increased expression of Stat3. Knockdown of PCNA, JNK1, or Stat3 did not have an effect on NPM and vice versa. In conclusion, our results suggest that PCNA and JNK1-Stat3 pathways respectively promotes and feedback inhibits the centrosome amplification by targeting at the ROCK1/14-3-3σ complex, and NPM serves as an independent signal for the centrosome amplification.© 2018 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

Keyword: colon cancer

and linoleic metabolism in Caco-2 cells: different triglyceride synthesis and lipoprotein secretion.

Polarized monolayers of intestinal Caco-2 cells were used to study the effects of saturated (16:0) and polyunsaturated linoleic (18:2) on triglyceride synthesis and lipoprotein secretion. Monolayers were incubated for 24 h, at the apical or lumenal side, with (16:0) or linoleic (18:2) in physiological concentrations. Incubation with 1.0 mM 16:0 or 18:2 resulted in differences in the composition and amount of secreted lipoproteins. Radiolabeled lipids in the lipoproteins secreted during incubation with 18:2 were found in the chylomicron/VLDL (very low density lipoprotein) density whereas with 16:0 the secreted lipoproteins were in the intermediate density/low density lipoprotein (IDL/LDL) density range. More triglyceride was secreted into the (basolateral) medium during incubation with 1.0 mM 18:2 (41 +/- 12% of total triglyceride synthesized) than with 1.0 mM 16:0 (18 +/- 3% of total). The biochemical findings correlate with conspicuous morphological changes in the cells in the presence of 16:0, but not 18:2. Increasing concentrations of 16:0 (0.1-1.0 mM) caused gradual accumulation of intracellular membrane. Microvilli became strongly reduced in number. With 1.0 mM we found an increased incorporation of [1-14C] into phosphatidic (14.8% of total incorporation into phospholipid with 16:0 vs. < 0.5% with 18:2) and diacylglycerol (12.5% with 16:0 vs. < 0.5% with 18:2) and diacylglycerol (12.5% with 16:0 vs. 0.5% with 18:2) and the amount of intracellular phospholipid doubled. The morphological changes were completely reversed after 24 h with 1.0 mM 18:2. We conclude from our results that, compared to 18:2, 16:0 is not efficiently incorporated into triglycerides. 16:0 is incorporated into cellular phospholipids in a greater proportion than 18:2, causing accumulation of intracellular phospholipid and the precursors phosphatidic and diacylglycerol. Different processing of 18:2 and 16:0 by Caco-2 cells resulted in profound differences in triglyceride synthesis and lipoprotein composition and secretion.

Keyword: colon cancer

Biochemical epidemiology of : effect of types of dietary fiber on diacylglycerols in women.

In view of the potential significance of dietary fat and fiber in and the possible indirect involvement of diacylglycerols (DAGs) in the pathogenesis of , the effect of types of dietary fiber on fecal DAG in premenopausal women was investigated.Forty-eight women consuming a typical western diet provided two 24-hour stool specimens and two sets of preintervention 4-day food records. They were randomly assigned to one of the fiber groups, namely, a wheat, oat, or corn bran supplement. They consumed their control diet plus 13-15 g of dietary fiber from each source for 8 weeks. At the end of the fiber period, each subject provided two 24-hour stool specimens and 4-day food records. Stool samples collected during the two periods were analyzed for total fat and DAG fatty acids.All sources of dietary fiber increased the amount of fecal fat excreted. Dietary wheat bran decreased the concentrations of total DAG and DAG containing lauric , myristic , , stearic , and linoleic , whereas oat bran increased the DAG composed of oleic and linoleic compared with the control diet. Corn bran decreased the DAG containing stearic .These results show that the modifying effect of dietary fiber on DAG depends on the type of fiber consumed.

Keyword: colon cancer

Modification of CaCo-2 cell membrane fatty composition by eicosapentaenoic and : effect on cholesterol metabolism.

Membrane fatty composition of CaCo-2 cells was modified by incubating the cells for 8 days in medium containing 100 microM eicosapentaenoic or . The effect of membrane fatty changes on cholesterol metabolism was then studied. Cells incubated with eicosapentaenoic had significant changes in membrane fatty composition with an accumulation of 20:5 and 22:5 and a reduction in monoenoic fatty acids compared to cells grown in . Intracellular cholesteryl esters could not be detected in CaCo-2 cells grown in the presence of the n-3 polyunsaturated fatty . In contrast, cells incubated with the saturated fatty contained 2 micrograms/mg protein of cholesteryl esters. Cells grown in eicosapentaenoic , however, accumulated significantly more triglycerides compared to cells modified with . The rate of oleic incorporation into triglycerides was significantly increased in cells incubated with eicosapentaenoic . CaCo-2 cells modified by eicosapentaenoic had lower rates of HMG-CoA reductase and ACAT activities compared to cells modified with . The incorporation of the two fatty acids into cellular lipids also differed. was predominantly incorporated into cellular triglycerides, whereas eicosapentaenoic was preferentially incorporated into phospholipids with 60% of it in the phosphatidylethanolamine fraction. The data indicate that membrane fatty composition is significantly altered by growing CaCo-2 cells in eicosapentaenoic . These modifications in membrane fatty saturation are accompanied by a decrease in the rates of cholesterol synthesis and cholesterol esterification.

Keyword: colon cancer

New flavan and alkyl alpha,beta-lactones from the stem bark of Horsfieldia superba.

In the present study phytochemical investigation of the methanol extract of the stem bark of Horsfieldia superba led to the isolation of twenty compounds (1-20), of which three (1-3) were new. However, compounds 2 and 3 were previously reported as synthetic alpha,beta-lactones. The compounds were characterized as (-)-3,4',7-trihydroxy-3'-methoxyflavan (1), (-)-5,6-dihydro-6-undecyl-2H-pyran-2-one (2), and (-)-5,6-dihydro-6-tridecyl-2H-pyran-2-one (3). Seventeen other known compounds were also isolated and identified as (-)-viridiflorol (4), hexacosanoic (5), beta-sitosterol (6), methyl 2,4-dihydroxy-6-methylbenzoate (methylorsellinate) (7), methyl 2,4-dihydroxy-3,6-dimethylbenzoate (8), (-)-4'-hydroxy-7-methoxyflavan (9), (-)-4',7-dihydroxyflavan (10), (-)-4',7-dihydroxy-3'-methoxyflavan (11), (+)-3,4',7-trihydroxyflavan (12), (-)-catechin (13), (-)-epicatechin (14), (-)-7-hydroxy-3',4'-methylenedioxyflavan (15), 2',3,4-trihydroxy-4'-methoxydihydrochalcone (16), 3',4',7-trihydroxyflavone (17), (+)-4'-hydroxy-7-methoxyflavanone (18), hexadecanoic () (19) and 3,4-dihydroxybenzoic (20). The structures of the compounds were fully characterized by various physical methods (melting point, optical rotation), spectral (UV, IR, ID and 2D NMR) and mass spectrometric techniques. In vitro assay of compounds 2 and 3 demonstrated moderate cytotoxic activities against human prostate (PC-3), (HCT-116) and breast (MCF-7) cells, while the chloroform and ethyl acetate fractions of H. superba were found to exhibit moderate AChE inhibitory activity (IC50 72 and 60 microg/mL).

Keyword: colon cancer

Altered Saturated and Monounsaturated Plasma Phospholipid Fatty Profiles in Adult Males with Adenomas.

Altered lipid metabolism and plasma fatty (FA) levels are associated with colorectal . Obesity and elevated waist circumference (WC) increase the likelihood of developing precancerous adenomas.Venous blood was collected from 126 males, ages 48 to 65 years, who received routine colonoscopies. Plasma phospholipid (PPL) FAs were isolated, derivatized, and then analyzed using gas chromatography. ORs and 95% confidence intervals were determined using polytomous logistic regression after adjusting for confounding factors [i.e., age, smoking, WC, and body mass index (BMI)].PPL (PA) was inversely correlated with the presence of adenomas (P = 0.01). For each unit increase in palmitoleic (OR, 3.75; P = 0.04) or elaidic (OR, 2.92; P = 0.04), an individual was more likely to have adenomas relative to no polyps. Higher enzyme activity estimates (EAE) of stearoyl-CoA desaturase-1 (SCD-1; P = 0.02) and elongation of very long chain fatty acids protein-6 (ELOVL-6; P = 0.03) were associated with an individual being approximately 1.5 times more likely to have an adenoma compared with no polyps.PPL FAs and EAEs, which have previously been associated with colorectal , are significantly different in those with adenomas when compared with those without polyps. PPL PA, elaidic , and SCD-1 and ELOVL-6 EAEs are associated with adenomas independent of BMI and WC.PPL PA, elaidic , and SCD-1 and ELOVL-6 EAEs are associated with adenomas even after adjusting for obesity-related risk factors and may function as novel biomarkers of early colorectal risk.©2015 American Association for Research.

Keyword: colon cancer

Non-proteolytic release of carcinoembryonic antigen from normal human epithelial cells cultured in collagen gel.

Recent studies have shown that, even with a minimal content of carcinoembryonic antigen (CEA), normal human epithelial cells express substantial amounts of CEA mRNA and mucosal fragments cultured in vitro produce CEA quite actively, indicating that CEA should no longer be considered to be of an oncofetal nature. To understand the basis of the usefulness of CEA as a tumor marker, we analyzed the release of CEA, a glycosyl-phosphatidylinositol (GPI)-anchored protein, from epithelial cells, by culturing isolated crypts in collagen gel. The crypts appeared to preserve their morphological and biochemical integrity in the gel for at least 16 hr, and released CEA spontaneously. Three forms of CEA--spontaneously released CEA, CEA liberated with phosphatidylinositol-specific phospholipase C (PI-PLC) and CEA in cell lysates--were indistinguishable on SDS-PAGE. This is in contrast to recombinant CEA spontaneously released from CHO transfectants, which showed a smaller molecular mass than that of PI-PLC-cleaved recombinant CEA. By phase separation using Triton X-114, CEA in the cell lysates of crypts was separated mostly into the detergent phase, while the spontaneously released and the PI-PLC-cleaved CEA were separated into the aqueous phase. When the cells were metabolically labeled with the precursors of the GPI-anchor, 3H-ethanolamine but not 3H- was found in the spontaneously released CEA. These findings suggest that, in contrast to the proteolysis-like release of the recombinant CEA from CHO cells, CEA in normal epithelial cells is released by a non-proteolytic cleavage, which probably occurs through the action of some endogenous phospholipase.

Keyword: colon cancer

Antitumor effect of -conjugated DsiRNA for in a mouse subcutaneous tumor model.

In this study, we synthesized Dicer-substrate siRNA conjugated with at the 5'-end of the sense strand (C16-DsiRNA), and examined its RNAi effect on β-catenin as a target gene in a cell line, HT29Luc, both in vitro and in vivo. We examined the in vitro RNAi effect in HT29Luc cells and found that C16-DsiRNA strongly inhibited expression of the β-catenin gene in comparison with non-modified DsiRNA. Also, high membrane permeability of C16-DsiRNA was exhibited, and it was confirmed that most of the C16-DsiRNA was localized in cytoplasm of HT29Luc cells. In regard to the in vivo RNAi effect, C16-DsiRNA complexed with Invivofectamine targeting the β-catenin gene was locally administered to a subcutaneous tumor formed by implantation of HT29Luc cells into the subcutis of nude mice; we evaluated the effect by measuring the bioluminescence increase, which reflects tumor growth, using an in vivo imaging system. As a result, C16-DsiRNA strongly inhibited the growth of tumors formed in subcutis of nude mice compared with non-modified DsiRNA, and this in vivo RNAi effect lasted up to 15\xa0days. Our results suggest that C16-DsiRNA should be vigorously pursued as a novel nucleic medicine for clinical treatment of .© 2018 John Wiley & Sons A/S.

Keyword: colon cancer

Urokinase secretion from human carcinomas induced by endogenous diglycerides.

tumor cells are more responsive to certain growth modulators in their local environment in vivo than are normal colonocytes. Examples of this class of compounds are the fecal diglycerides (DGs)(E. Friedman et al., Res., 49: 544-548, 1989), which may act as endogenous tumor promoters. At the concentration found in vivo, fecal DGs composed of oleic, myristic, and fatty acids induced mitogenesis of all classes of benign tumor cells and of half of the resected carcinomas tested in primary culture, but induced no detectable mitogenesis of normal colonocytes. tumor cells also exhibit selective responses to these endogenous modulators as measured by another biological parameter, secretion of urokinase from carcinomas than from normal colonocytes. Fecal DGs also induced a 13-fold increase in urokinase mRNA synthesis in carcinoma cells and induced secretion of active urokinase from each of five resected carcinomas. carcinomas, at both the primary site and metastatic to the liver, secreted the Mr 55,000 form of urokinase constitutively and secreted the same form upon treatment with fecal DGs. An increase in the steady-state level of urokinase secretion by saturated-chain DGs exhibited a strong dependency on the chain length of the fatty residues, those of 14 and 16 carbons having the greatest activity. Thus, fecal DGs composed of oleic, myristic, and residues induce two biological activities selectively in tumor cells, each of which would enhance tumor development. Selective mitogenesis would increase adenoma and carcinoma cell number relative to normal colonocyte number, and induction of the proteolytic enzyme urokinase would aid local invasion of the carcinoma within the bowel wall.

Keyword: colon cancer

Hexadecenoic Fatty Positional Isomers and De Novo PUFA Synthesis in Cells.

metabolism involves delta-9 and delta-6 desaturase enzymes forming palmitoleic (9-16:1; -7 series) and sapienic (6-16:1; -10 series), respectively. The corresponding biological consequences and lipidomic research on these positional monounsaturated fatty (MUFA) isomers are under development. Furthermore, sapienic can bring to the de novo synthesis of the -10 polyunsaturated fatty (PUFA) sebaleic (5,8-18:2), but such transformations in cells are not known. The model of Caco-2 cell line was used to monitor sapienic supplementation (150 and 300 μM) and provide evidence of the formation of -10 fatty acids as well as their incorporation at levels of membrane phospholipids and triglycerides. Comparison with palmitoleic and acids evidenced that lipid remodelling was influenced by the type of fatty and positional isomer, with an increase of 8-18:1, -10 PUFA and a decrease of saturated fats in case of sapienic . Cholesteryl esters were formed only in cases with sapienic . Sapienic was the less toxic among the tested fatty acids, showing the highest ECs and inducing death only in 75% of cells at the highest concentration tested. Two-photon fluorescent microscopy with Laurdan as a fluorescent dye provided information on membrane fluidity, highlighting that sapienic increases the distribution of fluid regions, probably connected with the formation of 8-18:1 and the -10 PUFA in cell lipidome. Our results bring evidence for MUFA positional isomers and de novo PUFA synthesis for developing lipidomic analysis and research.

Keyword: colon cancer

Incorporation of bile of low concentration into model and biological membranes studied by 2H and 31P NMR.

We have analyzed the manner of incorporation of bile into lipid bilayers and resultant perturbation of the bilayer structure with lower bile /lipid ratios relevant to the physiological conditions (approximately 1 mM) by 2H and 31P NMR methods, as an aid to understanding the possible role as an endogenous tumor promoter in besides the primary physiological function of solubilizing lipids. On the basis of the 2H quadrupole splittings of [6,6,7,7,8-2H5]deoxycholate and [11,11,12,12-2H4]chenodeoxycholate in the presence of lamellar multibilayers of egg yolk lecithin, these bile acids were found to be incorporated in such a manner that the B-D rings lie parallel with the normal of the bilayers when the ratio of the bile to lipid is low (less than 0.11). When the ratio is increased, these bile molecules are not dispersed entirely in the bilayer but aggregate to form micelles with lipids. Further, we studied the resultant perturbation of the multibilayers of egg yolk lecithin analyzed by using the 2H quadrupole splitting of [18,18,18-2H3]stearic as a probe and by 31P chemical shift anisotropy. We found that the bilayer structure is retained even at the bile -to-lipid ratio of 0.25, although a small amount of an isotropic phase appeared such as small vesicles and micelles. The molecular ordering of fatty acyl chains was rather enhanced by the presence of 1 mM deoxycholate in erythrocyte ghosts as seen from the 2H quadrupole splitting of [16,16,16-2H3], although deoxycholate caused hemolysis in this condition. The former observation can be explained by the way the lipid-protein interaction is modified by deoxycholate located in the interface between the lipids and proteins.

Keyword: colon cancer

Anti-Escherichia coli O157:H7 activity of free fatty acids under varying pH.

Following screening of 4 strains of Escherichia coli O157:H7 (E32511, E318N, H4420N, and R508N) for tolerance, strain H4420N was selected for further study into the influence of pH on bactericidal activity of 6 fatty acids (capric, lauric, , oleic, linoleic, and linolenic). Strain H4420N was cultured for 6 h in Luria-Bertani broth amended with individual fatty acids at 20 mmol/L, with pH adjusted to 7.0, 4.3, or 2.5. None of the fatty acids exhibited bactericidal activity at pH 7.0 (p >0.05). At pH 4.3, only capric, lauric, and linoleic acids reduced viability of H4420N (p < 0.05). At pH 2.5, oleic (C18:1) and linolenic (C18:3) acids had modest effects on H4420N viability, whereas capric (C10:0), lauric (C12:0), and linoleic (C18:2) acids resulted in a reduction > or =5 log10 colony-forming units (CFU)/mL (p < 0.05). Capric and lauric acids were examined further at pH 2.5 over a range of concentrations (0.15-20 mmol/L). After 10 min of exposure, 5 log10 CFU/mL reductions (p < 0.05) were achieved by lauric at 2.5 mmol/L and by capric at 0.31 mmol/L. stress increased the sensitivity of -tolerant E. coli O157:H7 strain H4420N to fatty acids. Including sources of these fatty acids in diets for cattle might impair the ability of this zoonotic pathogen to survive passage through the stomach, possibly reducing the potential for its in the lower gut.

Keyword: colonization

Metabolite profiling of mycorrhizal roots of Medicago truncatula.

Metabolite profiling of soluble primary and secondary metabolites, as well as cell wall-bound phenolic compounds from roots of barrel medic (Medicago truncatula) was carried out by GC-MS, HPLC and LC-MS. These analyses revealed a number of metabolic characteristics over 56 days of symbiotic interaction with the arbuscular mycorrhizal (AM) fungus Glomus intraradices, when compared to the controls, i.e. nonmycorrhizal roots supplied with low and high amounts of phosphate. During the most active stages of overall root mycorrhization, elevated levels of certain amino acids (Glu, Asp, Asn) were observed accompanied by increases in amounts of some fatty acids ( and oleic acids), indicating a mycorrhiza-specific activation of plastidial metabolism. In addition, some accumulating fungus-specific fatty acids (palmitvaccenic and vaccenic acids) were assigned that may be used as markers of fungal root . Stimulation of the biosynthesis of some constitutive isoflavonoids (daidzein, ononin and malonylononin) occurred, however, only at late stages of root mycorrhization. Increase of the levels of saponins correlated AM-independently with plant growth. Only in AM roots was the accumulation of apocarotenoids (cyclohexenone and mycorradicin derivatives) observed. The structures of the unknown cyclohexenone derivatives were identified by spectroscopic methods as glucosides of blumenol C and 13-hydroxyblumenol C and their corresponding malonyl conjugates. During mycorrhization, the levels of typical cell wall-bound phenolics (e.g. 4-hydroxybenzaldehyde, vanillin, ferulic ) did not change; however, high amounts of cell wall-bound tyrosol were exclusively detected in AM roots. Principal component analyses of nonpolar primary and secondary metabolites clearly separated AM roots from those of the controls, which was confirmed by an hierarchical cluster analysis. Circular networks of primary nonpolar metabolites showed stronger and more frequent correlations between metabolites in the mycorrhizal roots. The same trend, but to a lesser extent, was observed in nonmycorrhizal roots supplied with high amounts of phosphate. These results indicate a tighter control of primary metabolism in AM roots compared to control plants. Network correlation analyses revealed distinct clusters of amino acids and sugars/aliphatic acids with strong metabolic correlations among one another in all plants analyzed; however, mycorrhizal symbiosis reduced the cluster separation and enlarged the sugar cluster size. The amino clusters represent groups of metabolites with strong correlations among one another (cliques) that are differently composed in mycorrhizal and nonmycorrhizal roots. In conclusion, the present work shows for the first time that there are clear differences in development- and symbiosis-dependent primary and secondary metabolism of M. truncatula roots.

Keyword: colonization

Deficient production of hexadecenoic in the skin is associated in part with the vulnerability of atopic dermatitis patients to by Staphylococcus aureus.

As one of the major skin fatty acids, cis-6-hexadecenoic (C16:1Delta6) exhibits a specific antibacterial activity and might play a specific role in the defense mechanism against Staphylococcus aureus, in healthy subjects whereas S. aureus frequently colonizes the skin of patients with atopic dermatitis (AD).Fatty composition of sebum at the recovery level was analyzed by gas chromatography and S. aureus colonizing the skin was assessed by the \'cup-scrub\' method (9 patients and 10 healthy controls). To evaluate in vivo effect of C16:1Delta6 on , C16:1Delta6 was applied for 2 weeks on the upper arm skin of another group of AD patients (11 patients).Analysis of sebum lipids revealed that there is a significant lower free C16:1Delta6 content in nonlesional skin from AD patients compared with healthy controls. This lower content is also accompanied by a significantly lower level of C16:1Delta6 in the total fatty composition of sebum (analyzed following hydrolysis). The lower level of free C16:1Delta6 correlated significantly (R(2) = 0.41, p < 0.01) with the numbers of S. aureus colonizing nonlesional skin. Topical application of free C16:1Delta6 on the skin of AD patients for 2 weeks abolished the markedly increased bacterial count in 6 out of the 8 AD patients tested.Free C16:1Delta6 may be involved in the defense mechanism against S. aureus in healthy skin and this deficit triggers the susceptibility of the skin to by S. aureus in AD.Copyright 2005 S. Karger AG, Basel.

Keyword: colonization

Human antibody response to outer membrane protein G1a, a lipoprotein of Moraxella catarrhalis.

Moraxella catarrhalis is an important cause of respiratory infections in adults with chronic obstructive pulmonary disease (COPD) and of otitis media in children. Outer membrane protein (OMP) G1a is an approximately 29-kDa surface lipoprotein and is a potential vaccine candidate. The gene that encodes OMP G1a was expressed and purified using a novel plasmid vector. [(3)H] labeling demonstrated that both native and recombinant OMP G1a contain covalently bound . To assess the expression of OMP G1a during human infection, paired sera and sputum supernatants from adults with COPD followed prospectively were studied by enzyme-linked immunosorbent assays with recombinant lipidated OMP G1a to detect antibodies made specifically during carriage of M. catarrhalis. Overall, 23% of patients developed either a serum immunoglobulin G (IgG) response (9%) or sputum IgA response (21%) to OMP G1a, following 100 episodes of acquisition and clearance of M. catarrhalis. Patients developed antibody responses at similar rates following episodes of clinical exacerbation compared to asymptomatic . Serum IgG antibodies following natural infection were directed predominantly at OMP G1a epitopes that are not exposed on the bacterial surface. These data show that OMP G1a is expressed during infection of the human respiratory tract and is a target of systemic and mucosal antibodies. These observations indicate that OMP G1a, a highly conserved surface protein, should be evaluated further as a vaccine candidate.

Keyword: colonization

In vitro and in vivo exploration of from Synechococcus elongatus as an antibiofilm agent on the survival of Artemia franciscana against virulent vibrios.

Biofilm formation of Vibrio spp. has been demonstrated as a potentially important mechanism contributing antibiotic treatment failure in aquaculture. In the present study, the effect of (PA) identified from Synechococcus elongatus was assessed for the inhibition of quorum sensing (QS) regulated biofilm formation in aquatic bacterial pathogens. The biofilm inhibitory concentration (BIC) of PA against Vibrio spp. was found to be 100µgml. In this concentration, PA exhibited a significant inhibition in biofilm biomass of Vibrio harveyi MTCC 3438, V. parahaemolyticus ATCC 17802, V. vulnificus MTCC 1145 and V. alginolyticus ATCC 17749 without hindering their planktonic growth. Also, PA displayed gradual decrease in bioluminescence production of V. harveyi. The results of extracellular polymeric substances quantification, microbial adhesion to hydrocarbons and Fourier transform infrared spectroscopic (FT-IR) analyses suggested that PA positively interferes with the initial adhesion stages of biofilm formation. In addition, confocal and scanning electron microscopic analysis substantiates the antibiofilm efficacy of the PA. The transcriptomic analysis revealed the down-regulation of QS mediated response regulator genes expression in V. harveyi. Concomitantly, PA reduced the intestinal of vibrios in brine shrimp larvae and thereby attenuates the biofilm assemblage and its associated virulence. In vivo studies using brine shrimp larvae manifested the reduction in adherence and virulence, which prompts further investigation about the potential of PA for the treatment of vibriosis.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: colonization

Enterobacter sp. strain Fs-11 adapted to diverse ecological conditions and promoted sunflower achene yield, nutrient uptake, and oil contents.

Plant growth-promoting rhizobacteria are under extensive investigation to supplement the chemical fertilizers due to cost-effective and eco-friendly nature. However, their consistency in heterogeneous soil and diverse ecological settings is unclear. The current study presents in vitro and field evaluation of pre-characterized PGPR strain Enterobacter sp. Fs-11 (GenBank accession # GQ179978) in terms of its potential to enhance sunflower yield and oil contents under diverse environmental conditions. Under in vitro conditions, strain Fs-11 showed optimal growth at a range of temperature (15 to 40\xa0°C) and pH values (6.5 to 8.5). Extracellular and intracellular localizations of the strain Fs-11 in sunflower root cortical cells through transmission electron microscopy confirmed its epiphytic and endophytic patterns, respectively. In field experiments, conducted at three different agro-climatic locations, inoculation of strain Fs-11 at 50% reduced NP fertilizer resulted in a significant increase in growth, achene yield, nutrient uptake, and oil contents. Inoculation also responded significantly in terms of increase in mono- and polyunsaturated fatty acids (oleic and linoleic acids, respectively) without rising saturated fatty ( and stearic acids) contents. We concluded that Enterobacter sp. Fs-11 is a potential candidate for biofertilizer formulations to supplement chemical fertilizer requirements of sunflower crop under diverse climatic conditions.

Keyword: colonization

Hydrophobicity of cultured mammalian cells and some effects of bacterial phospholipases C.

Eucaryotic cell surface hydrophobicity was measured as a partition of between the cell surface and the surrounding buffer. The method was found to be independent of cell mass or amount of within a large interval. An estimation of cell stability could also be obtained. The effects of Ca and Mg ions on cell hydrophobicity and stability of mouse myeloma (SP2/O) cells and of Chinese hamster ovary (CHO) cells were determined. This system permits measurement of changes in cell hydrophobicity caused by various additives, e.g. ions, purified bacterial products, antibiotics or cytostatics. Studies were made on these eucaryotic cells treated with purified bacterial phospholipases C from S. aureus and C. perfringens. These enzymes were found to increase the eucaryotic cell membrane hydrophobicity. This finding might indicate that bacterial phospholipases C facilitate bacterial at the start of an infection.

Keyword: colonization

Bactericidal activity of the human skin fatty cis-6-hexadecanoic on Staphylococcus aureus.

Human skin fatty acids are a potent aspect of our innate defenses, giving surface protection against potentially invasive organisms. They provide an important parameter in determining the ecology of the skin microflora, and alterations can lead to increased by pathogens such as Staphylococcus aureus. Harnessing skin fatty acids may also give a new avenue of exploration in the generation of control measures against drug-resistant organisms. Despite their importance, the mechanism(s) whereby skin fatty acids kill bacteria has remained largely elusive. Here, we describe an analysis of the bactericidal effects of the major human skin fatty cis-6-hexadecenoic (C6H) on the human commensal and pathogen S. aureus. Several C6H concentration-dependent mechanisms were found. At high concentrations, C6H swiftly kills cells associated with a general loss of membrane integrity. However, C6H still kills at lower concentrations, acting through disruption of the proton motive force, an increase in membrane fluidity, and its effects on electron transfer. The design of analogues with altered bactericidal effects has begun to determine the structural constraints on activity and paves the way for the rational design of new antistaphylococcal agents.Copyright © 2014 Cartron et al.

Keyword: colonization

Lipid metabolism in arbuscular mycorrhizal roots of Medicago truncatula.

The peroxidation of polyunsaturated fatty acids, common to all eukaryotes, is mostly catalyzed by members of the lipoxygenase enzyme family of non-heme iron containing dioxygenases. Lipoxygenase products can be metabolized further in the oxylipin pathway by several groups of CYP74 enzymes. One prominent oxylipin is jasmonic (JA), a product of the 13-allene oxide synthase branch of the pathway and known as signaling substance that plays a role in vegetative and propagative plant development as well as in plant responses to wounding and pathogen attack. In barley roots, JA level increases upon by arbuscular mycorrhizal fungi. Apart from this first result regarding JA, no information is available on the relevance of lipidperoxide metabolism in arbuscular mycorrhizal symbiosis. Thus we analyzed fatty and lipidperoxide patterns in roots of Medicago truncatula during mycorrhizal . Levels of fungus-specific fatty acids as well as (16:0) and oleic (18:1 n - 9) were increased in mycorrhizal roots. Thus the degree of arbuscular mycorrhizal of roots can be estimated via analysis of fungal specific esterified fatty acids. Otherwise, no significant changes were found in the profiles of esterified and free fatty acids. The 9- and 13-LOX products of linoleic and alpha-linolenic were present in all root samples, but did not show significant differences between mycorrhizal and non-mycorrhizal roots, except JA which showed elevated levels in mycorrhizal roots. In both types of roots levels of 13-LOX products were higher than those of 9-LOX products. In addition, three cDNAs encoding CYP74 enzymes, two 9/13-hydroperoxide lyases and a 13-allene oxide synthase, were isolated and characterized. The transcript accumulation of these three genes, however, was not increased in mycorrhizal roots of M. truncatula.

Keyword: colonization

Phytoremediation assisted by mycorrhizal fungi of a Mexican defunct lead- battery recycling site.

A field experiment was conducted during 15\u202fmonths to study the effects of four arbuscular mycorrhizal fungi (AMF) on the growth of Ricinus communis accession SF7. Plants were established on amended soil (vermicompost:sawdust:soil 1:1:1) severely polluted by lead- batteries (LAB) located at Mexico State, Mexico. Plants inoculated with Acaulospora sp., Funneliformis mosseae and Gigaspora gigantea had 100% survival in comparison to non-inoculated plants (57%). These same AMF enhanced and linoleic acids content in seeds of R. communis. Acaulospora sp. modified rhizosphere soil pH and decreased 3.5 folds Pb foliar concentrations while F. mosseae BEG25 decreased three times Pb soil availability in comparison to non-inoculated plants. Spatial changes in Pb soil availability were observed at the end of this research. No fungal effect on P, Ca, Cu foliar concentrations, soluble sugars, proline, chlorophyll or on the activity of two oxidative stress enzymes was observed. Mycorrhizal from the inoculated fungi was between 40% and 60%, while by native fungi was between 16% and 22%. A similar percentage of foliar total phenolic compounds was observed in non-mycorrhizal plants and those inoculated with G. gigantea and Acaulospora sp. This is the first research reporting effects of AMF on R. communis (castor bean) shrubs when grown on a LAB recycling site suggesting the use of Acaulospora sp. and F. mosseae BEG25 in phytostabilization to ameliorate Pb pollution and decreasing its ecological risk.Copyright © 2018. Published by Elsevier B.V.

Keyword: colonization

New anti-infective coatings of surgical sutures based on a combination of antiseptics and fatty acids.

Wound infection is a complication feared in surgery. The aim of this study was to develop new anti-infective coatings of surgical sutures and to compare the anti-microbial effectiveness and biocompatibility to the well-established Vicryl Plus. Synthetic absorbable PGA surgical sutures were coated with three different chlorhexidine concentrations and two different octenidine concentrations in combination with and lauric . Drug-release kinetics lasting 96 h were studied in phosphate-buffered saline at 37 degrees C. Anti-infective characteristics were determined by measuring the change in optical density of Staphylococcus aureus suspensions charged with coated sutures over time. Microorganisms adsorbed at the surface of coated sutures were assessed on blood agar plates and coated sutures eluted for 24 h were placed on bacterial lawns cultured on Mueller-Hinton plates to prove retained anti-microbial potency. A cell proliferation assay was performed to assess the degree of cytotoxicity. Anti-infective characteristics and biocompatibility were compared to Vicryl Plus. A coating technology for slow-release drug-delivery systems on surgical sutures could be developed. All coatings showed a continuous drug release within 96 h. Individual chlorhexidine and octenidine coated sutures showed superior anti-infective characteristics but inferior biocompatibility in comparison to Vicryl Plus. We conclude that the developed anti-infective suture coatings consisting of lipid-based drug-delivery systems in combination with antiseptics are highly effective against bacterial in vitro; however, drug doses have to be adjusted to improve biocompatibility.

Keyword: colonization

ToxR regulates the production of lipoproteins and the expression of serum resistance in Vibrio cholerae.

The genes encoding three lipoproteins of Vibrio cholerae were identified by a combination of DNA sequence analysis and [3H]palmitate labeling of hybrid proteins encoded by TnphoA gene fusions. The expression of these three lipoproteins, TagA, AcfD, and TcpC, was controlled by ToxR, the cholera toxin transcriptional activator. The involvement of other bacterial lipoproteins in conferring resistance to the bactericidal effects of complement prompted us to examine this possibility in V. cholerae. Remarkably, mutations in toxR and tcp genes (including tcpC), involved in the biogenesis of the toxin coregulated pili, rendered V. cholerae about 10(4)-10(6) times more sensitive to the vibriocidal activity of antibody and complement. Since V. cholerae is a noninvasive organism and toxR and tcp mutants are highly defective in intestinal in animals and humans, these results raise the possibility that resistance to a gut-associated, "complement-like" bactericidal activity may be a major virulence determinant of V. cholerae and other enterobacterial species.

Keyword: colonization

Cell membrane-binding properties of group A streptococcal lipoteichoic .

Lipoteichoic (LTA) was extracted from group A streptococci, previously treated with hot HCl, by the phenol method. The extracted LTA was loaded on an isoelectric (IE) focusing column and two fractions were collected; one at pH 4.65 and the other at pH 2.95. Chemical analysis demonstrated that the unfractionated LTA contained alanine and glycerolphosphate at molar ratio of 1:10, and ester-linked lipids, but no detectable sugars or amino-sugars. The two IE fractions contained lipids but lacked alanine. The LTA and its IE fractions spontaneously adsorbed to human erythrocytes (sensitization) causing them to agglutinate in the presence of rabbit anti-LTA. The RBC-sensitizing and antigenic activities of IE fractions were equal to, or greater (for IE fraction at pH 4.65) than the unfractionated LTA, indicating that alanine is not involved in the sensitizing activity of LTA. Mild ammonia-hydrolysis abolished the RBC-sensitizing activity of LTA and its IE fractions. Chloroform-methanol-soluble material of the ammonia-hydrolysate lacked antigenic activity but blocked sensitization of erythrocytes by LTA. The water-soluble material of the hydrolyzed LTA retained antigenic activity, was not able to block sensitization by LTA, and its sensitizing activity was restored after esterification with fatty acids. These experiments indicate that ester-linked fatty acids ( being the major one) are involved in the spontaneous adsorption of LTA to erythrocytes. The LTA, its lipid moiety, and anti-LTA blocked adherence of group A streptococci to human epithelial cells, suggesting that small amounts of LTA may reside on the streptococcal surface to mediate attachment and of these organisms on mucosal surfaces in vivo.

Keyword: colonization

Adherence, coaggregation, and hydrophobicity of Streptococcus gordonii associated with expression of cell surface lipoproteins.

Streptococcus gordonii Challis incorporated exogenous [3H]palmitate into 13 polypeptides extractable from intact cells with sodium dodecyl sulfate. A 76-kDa surface-exposed polypeptide, implicated previously as a cell aggregation determinant, was shown to be one of these lipid-modified polypeptides. Differences in sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of lipopolypeptides were detected with mutants of S. gordonii that were altered in adherence, aggregation, coaggregation, or hydrophobicity. Lipid-modified polypeptides, tightly associated with the cell membrane, may be involved in the expression of cell surface properties of S. gordonii important for of the human oral cavity.

Keyword: colonization

Antiadhesive activity of the biosurfactant pseudofactin II secreted by the Arctic bacterium Pseudomonas fluorescens BD5.

Pseudofactin II is a recently identified biosurfactant secreted by Pseudomonas fluorescens BD5, the strain obtained from freshwater from the Arctic Archipelago of Svalbard. Pseudofactin II is a novel compound identified as cyclic lipopeptide with a connected to the terminal amino group of eighth amino in peptide moiety. The C-terminal carboxylic group of the last amino forms a lactone with the hydroxyl of Thr3. Adhesion is the first stage of biofilm formation and the best moment for the action of antiadhesive and anti-biofilm compounds. Adsorption of biosurfactants to a surface e.g. glass, polystyrene, silicone modifies its hydrophobicity, interfering with the microbial adhesion and desorption processes. In this study the role and applications of pseudofactin II as a antiadhesive compound has been investigated from medicinal and therapeutic perspectives.Pseudofactin II lowered the adhesion to three types of surfaces (glass, polystyrene and silicone) of bacterial strains of five species: Escherichia coli, Enterococcus faecalis, Enterococcus hirae, Staphylococcus epidermidis, Proteus mirabilis and two Candida albicans strains. Pretreatment of a polystyrene surface with 0.5 mg/ml pseudofactin II inhibited bacterial adhesion by 36-90% and that of C. albicans by 92-99%. The same concentration of pseudofactin II dislodged 26-70% of preexisting biofilms grown on previously untreated surfaces. Pseudofactin II also caused a marked inhibition of the initial adhesion of E. faecalis, E. coli, E. hirae and C. albicans strains to silicone urethral catheters. The highest concentration tested (0.5 mg/ml) caused a total growth inhibition of S. epidermidis, partial (18-37%) inhibition of other bacteria and 8-9% inhibition of C. albicans growth.Pseudofactin II showed antiadhesive activity against several pathogenic microorganisms which are potential biofilm formers on catheters, implants and internal prostheses. Up to 99% prevention could be achieved by 0.5 mg/ml pseudofactin II. In addition, pseudofactin II dispersed preformed biofilms. Pseudofactin II can be used as a disinfectant or surface coating agent against microbial of different surfaces, e.g. implants or urethral catheters.

Keyword: colonization

and mineralization of byPseudomonas pseudoflava.

Pseudomonas pseudoflava and were used to investigate the role of bacterial in the degradation of waterinsoluble organic compounds. Mineralization was measured by trapping the(14)CO2 produced from the labeled substrate, and of the surface of the solid organic chemical was determined by epifluorescence microscopy. In a medium containing solid ,P. pseudoflava mineralized the organic substrate at a logarithmic rate. Mineralization was evident before of the surface of the chemical was detected. The rate of appearance of single cells and/or aggregations of cells on the surface of the was essentially the same as the doubling time of free cells in solution. At about 50 hours, mineralization and of the surface stopped. In a salts solution containing solid ,P. pseudoflava grew logarithmically in the solution and biphasically on the surface of the . We suggest that the bacterium first metabolizes soluble and later colonizes the solid when the substrate in solution has been depleted.

Keyword: colonization

Docosahexaenoic and Eicosapentaenoic Acids Prevent Altered-Muc2 Secretion Induced by by Alleviating Endoplasmic Reticulum Stress in LS174T Goblet Cells.

Diets high in saturated fatty acids (FA) represent a risk factor for the development of obesity and associated metabolic disorders, partly through their impact on the epithelial cell barrier integrity. We hypothesized that unsaturated FA could alleviate saturated FA-induced endoplasmic reticulum (ER) stress occurring in intestinal secretory goblet cells, and consequently the reduced synthesis and secretion of mucins that form the protective mucus barrier. To investigate this hypothesis, we treated well-differentiated human colonic LS174T goblet cells with (PAL)-the most commonly used inducer of lipotoxicity in in vitro systems-or -9, -6, or -3 unsaturated fatty acids alone or in co-treatment with PAL, and measured the impact of such treatments on ER stress and Muc2 production. Our results showed that only eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids protect goblet cells against ER stress-mediated altered Muc2 secretion induced by PAL, whereas neither linolenic nor -9 and -6 FA are able to provide such protection. We conclude that EPA and DHA could represent potential therapeutic nutrients against the detrimental lipotoxicity of saturated fatty acids, associated with type 2 and obesity or inflammatory bowel disease. These in vitro data remain to be explored in vivo in a context of dietary obesity.

Keyword: diabetes

Irisin Ameliorates Glucolipotoxicity-Associated β-Cell Dysfunction and Apoptosis via AMPK Signaling and Anti-Inflammatory Actions.

Islet metabolic disorder and inflammation contribute to the pathogenesis and progression of type 2 (T2DM). Irisin is a recently identified adipomyokine with protective effects on metabolic homeostasis and inflammation-suppressing effects in hepatic and vascular cells. The present study examined the effects of irisin on lipid metabolism and inflammation in β cells under glucolipotoxic conditions.Rat INS-1E β cells and islets isolated from C57BL/6 mice were incubated in glucolipotoxic conditions with or without irisin. Intracellular lipid contents and lipogenic gene expression were determined by enzymatic colorimetric assays and real-time PCR, respectively. Inflammatory status was evidenced by Western blot analysis for the phosphorylation of nuclear factor-κB (NF-κB) p65 and real-time PCR analysis for the expression of pro-inflammatory genes.Irisin reversed glucolipotoxicity-induced intracellular non-esterified fatty (NEFA) and triglyceride accumulation, suppressed associated elevations in lipogenic gene expression, and phosphorylated acetyl-CoA-carboxylase (ACC) in INS-1E cells. These demonstrated effects were dependent on irisin-activated adenosine monophosphate-activated protein kinase (AMPK). Meanwhile, AMPK signaling mediated the protective effects of irisin on INS-1E cell insulin secretory ability and survival as well. Additionally, irisin inhibited phosphorylation of NF-κB p65 while decreasing the expression of pro-inflammatory genes in INS-1E cells under glucolipotoxic conditions. Moreover, irisin also improved insulin secretion, inhibited apoptosis, and restored β-cell function-related gene expression in isolated mouse islets under glucolipotoxic conditions.Irisin attenuated excessive lipogenesis in INS-1E cells under glucolipotoxic state through activation of AMPK. Irisin also suppressed overnutrition-induced inflammation in INS-1E cells. Our findings implicate irisin as a promising therapeutic target for the treatment of islet lipid metabolic disorder and islet inflammation in T2DM.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

BH3 mimetics derived from Bim-BH3 domain core region show PTP1B inhibitory activity.

A series of our previously described BH3 peptide mimetics derived from Bim-BH3 domain core region were found to exhibit weak to potent PTP1B binding affinity and inhibitory activities via target-based drug screening. Among these compounds, a 12-aa Bim-BH3 core sequence peptide conjugated to (SM-6) displayed good PTP1B binding affinity (K\u202f=\u202f8.38\u202fnmol/L), inhibitory activity (IC\u202f=\u202f1.20\u202fμmol/L) and selectivity against other PTPs (TCPTP, LAR, SHP-1 and SHP-2). Furthermore, SM-6 promoted HepG2 cell glucose uptake and inhibited the expression of PTP1B, indicating that SM-6 could improve the insulin resistance effect in the insulin-resistant HepG2 cell model. These results may indicate a new direction for the application of BH3 peptide mimetics and promising PTP1B peptide inhibitors could be designed and developed based on SM-6.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Characterization of α-Glucosidase Inhibitors from Lindau Leaves by Gas Chromatography-Mass Spectrometry-Based Metabolomics and Molecular Docking Simulation.

() is an herbal shrub traditionally consumed to treat various diseases including in Malaysia. This study was designed to evaluate the α-glucosidase inhibitory activity of leaves extracts, and to identify the metabolites responsible for the bioactivity.Crude extract obtained from the dried leaves using 80% methanolic solution was further partitioned using different polarity solvents. The resultant extracts were investigated for their α-glucosidase inhibitory potential followed by metabolites profiling using the gas chromatography tandem with mass spectrometry (GC-MS).Multivariate data analysis was developed by correlating the bioactivity, and GC-MS data generated a suitable partial least square (PLS) model resulting in 11 bioactive compounds, namely, , phytol, hexadecanoic (methyl ester), 1-monopalmitin, stigmast-5-ene, pentadecanoic , heptadecanoic , 1-linolenoylglycerol, glycerol monostearate, alpha-tocospiro B, and stigmasterol. In-silico study via molecular docking was carried out using the crystal structure isomaltase (PDB code: 3A4A). Interactions between the inhibitors and the protein were predicted involving residues, namely LYS156, THR310, PRO312, LEU313, GLU411, and ASN415 with hydrogen bond, while PHE314 and ARG315 with hydrophobic bonding.The study provides informative data on the potential α-glucosidase inhibitors identified in leaves, indicating the plant\'s therapeutic effect to manage hyperglycemia.

Keyword: diabetes

Scopoletin intervention in pancreatic endoplasmic reticulum stress induced by lipotoxicity.

Endoplasmic reticulum (ER), a dynamic organelle, plays an essential role in organizing the signaling pathways involved in cellular adaptation, resilience, and survival. Impairment in the functions of ER occurs in a variety of nutritive disorders including obesity and type 2 . Here, we hypothesize that (scopoletin) SPL, a coumarin, has the potential to alleviate ER stress induced in vitro and in vivo models by lipotoxicity. To test this hypothesis, the ability of SPL to restore the levels of proteins of ER stress was analyzed. Rat insulinoma 5f (RIN5f) cells and Sprague Dawley rats were the models used for this study. Groups of control and high-fat, high-fructose diet (HFFD)-fed rats were treated with either SPL or 4-phenylbutyric . Status of ER stress was enumerated by quantitative RT-PCR, Western blot, electron microscopic, and immunohistochemical studies. Proximal proteins of ER stress inositol requiring enzyme 1 (IRE1), protein kinase like endoplasmic reticulum kinase (PERK), and activating transcription factor 6 (ATF6) were reduced in the β-cells by SPL. The subsequent signaling proteins X-box binding protein 1, eukaryotic initiation factor2α, activating transcription factor 4, and C/EBP homologous protein were also suppressed in their expression levels when treated with SPL. IRE1, PERK signaling leads to c-Jun-N-terminal kinases phosphorylation, a kinase that interrupts insulin signaling, which was also reverted upon scopoletin treatment. Finally, we confirm that SPL has the ability to suppress the stress proteins and limit pancreatic ER stress which might help in delaying the progression of insulin resistance.

Keyword: diabetes

The Mitochondria-Targeted Antioxidant MitoQ Modulates Mitochondrial Function and Endoplasmic Reticulum Stress in Pancreatic β Cells Exposed to Hyperglycaemia.

Mitochondria-targeted antioxidants such as mitoquinone (MitoQ) have demonstrated protective effects against oxidative damage in several diseases. The increase in reactive oxygen species (ROS) production during glucose metabolism in β cells can be exacerbated under hyperglycaemic conditions such as type 2 (T2D), thus contributing to β cell function impairment. In the present work, we aimed to evaluate the effect of MitoQ on insulin secretion, oxidative stress, endoplasmic reticulum (ER) stress and nuclear factor kappa B (NFκB) signalling in a pancreatic β cell line under normoglycaemic (NG, 11.1 mM glucose), hyperglycaemic (HG, 25 mM glucose) and lipidic ( (PA), 0.5mM) conditions.We incubated the pancreatic β cell line INS-1E with or without MitoQ (0.5µM) under NG, HG and PA conditions. We then assessed the following parameters: glucose-induced insulin secretion, O₂ consumption (with a Clark-type electrode); mitochondrial function, oxidative stress parameters and calcium levels (by fluorescence microscopy); ER stress markers and NFκB-p65 protein levels (by western blotting).MitoQ increased insulin secretion and prevented the enhancement of ROS production and O₂ consumption and decrease in GSH levels that are characteristic under HG conditions. MitoQ also reduced protein levels of ER stress markers (GRP78 and P-eIF2α) and the proinflammatory nuclear transcription factor NFκB-p65, both of which increased under HG. MitoQ did not significantly alter ER stress markers under lipidic conditions.Our findings suggest that treatment with MitoQ modulates mitochondrial function, which in turn ameliorates endoplasmic reticulum stress and NFκB activation, thereby representing potential benefits for pancreatic β cell function.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: diabetes

Palmitate is not an effective fuel for pancreatic islets and amplifies insulin secretion independent of calcium release from endoplasmic reticulum.

The aim of the study was to determine the acute contribution of fuel oxidation in mediating the increase in insulin secretion rate (ISR) in response to fatty acids. Measures of mitochondrial metabolism, as reflected by oxygen consumption rate (OCR) and cytochrome c reduction, calcium signaling, and ISR by rat islets were used to evaluate processes stimulated by acute exposure to (PA). The contribution of mitochondrial oxidation of PA was determined in the presence and absence of a blocker of mitochondrial transport of fatty acids (etomoxir) at different glucose concentrations. Subsequent to increasing glucose from 3 to 20 mM, PA caused small increases in OCR and cytosolic calcium (about 20% of the effect of glucose). In contrast, the effect of PA on ISR was almost 3 times that by glucose, suggesting that the metabolism of PA is not the dominant mechanism mediating PA\'s effect on ISR. This was further supported by lack of inhibition of PA-stimulated OCR and ISR when blocking entry of PA into mitochondria (with etomoxir), and PA\'s lack of stimulation of reduced cytochrome c in the presence of high glucose. Consistent with the lack of metabolic stimulation by PA, an inhibitor of calcium release from the endoplasmic reticulum, but not a blocker of L-type calcium channels, abolished the PA-induced elevation of cytosolic calcium. Notably, ISR was unaffected by thapsigargin showing the dissociation of endoplasmic reticulum calcium release and second phase insulin secretion. In conclusion, stimulation of ISR by PA was mediated by mechanisms largely independent of the oxidation of the fuel.

Keyword: diabetes

Docosahexaenoic Reduces -Induced Endoplasmic Reticulum Stress in Pancreatic Β Cells.

Endoplasmic reticulum (ER) stress leads to peripheral insulin resistance and the progression of pancreatic beta cell failure in type 2 . Although ER stress plays an important role in the pathogenesis of , it is indispensable for cellular activity. Therefore, when assessing the pathological significance of ER stress, it is important to monitor and quantify ER stress levels. Here, we have established a novel system to monitor ER stress levels quickly and sensitively, and using this method, we have clarified the effect of differences in glucose concentration and various fatty acids on the ER of pancreatic β cells. First, we developed a cell system that secretes Gaussia luciferase in culture medium depending on the activation of the GRP78 promoter. This system could sensitively monitor ER stress levels that could not be detected with real-time RT-PCR and immunoblotting. This system revealed that hyperglycemia does not induce unfolded protein response (UPR) in a short period of time in MIN6 cells, a mouse pancreatic β cell line. Physiological concentrations of , a saturated fatty , induced ER stress quickly, while physiological concentrations of oleic , an unsaturated fatty , did not. Docosahexaenoic , an n-3 unsaturated fatty , inhibited -induced ER stress. In this study, we have established a system that can sensitively detect ER stress levels of living cells in a short period of time. This system can be used to monitor the state of the ER in living cells and lead to the investigation of the significance of physiological or pathological ER stress levels.

Keyword: diabetes

The protective role of the MKP-5-JNK/P38 pathway in glucolipotoxicity-induced islet β-cell dysfunction and apoptosis.

Hyperglycemia and hyperlipidemia (glycolipotoxicity)-triggered islet β-cell dysfunction is known to drive the progression of obesity-related type 2 , however the underlying mechanisms have not been clearly elucidated. The current study aimed to investigate the role of mitogen-activated protein kinase phosphatase 5 (MKP-5) in islet cells under glucolipotoxic conditions. Using gene overexpression and knockdown approaches, we demonstrated that MKP-5 could alleviate glucolipotoxicity-induced apoptosis via the endoplasmic reticulum (ER) stress and mitochondrial apoptosis pathways owing to the altered regulation of caspase family members and ER stress-related molecules in MIN6 and primary islet cells. Overexpression of MKP-5 reversed the glucose and (GP)-induced impairment of insulin secretion as well as the abnormal decreases in the expression of islet functional genes, thereby maintaining the normal insulin secretory functionality, whereas the absence of MKP-5 aggravated islet cell dysfunction. In parallel, the production of ROS and increased inflammation-associated genes in response to GP were also reduced upon MKP-5 overexpression. Further, inhibition of JNK or P38 MAPK pathways resisted to glucolipotoxicity observed in MKP-5 knockdown MIN6 cells. These findings indicate that MKP-5 is an important mediator for glucolipotoxicity-induced islet cell dysfunction and apoptosis, with JNK and P38 as the critical downstream pathways.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Probing the Global Cellular Responses to Lipotoxicity Caused by Saturated Fatty Acids.

Excessive levels of saturated fatty acids are toxic to cells, although the basis for this lipotoxicity remains incompletely understood. Here, we analyzed the transcriptome, lipidome, and genetic interactions of human leukemia cells exposed to palmitate. Palmitate treatment increased saturated glycerolipids, accompanied by a transcriptional stress response, including upregulation of the endoplasmic reticulum (ER) stress response. A comprehensive genome-wide short hairpin RNA (shRNA) screen identified >350 genes modulating lipotoxicity. Among previously unknown genetic modifiers of lipotoxicity, depletion of RNF213, a putative ubiquitin ligase mutated in Moyamoya vascular disease, protected cells from lipotoxicity. On a broader level, integration of our comprehensive datasets revealed that changes in di-saturated glycerolipids, but not other lipid classes, are central to lipotoxicity in this model. Consistent with this, inhibition of ER-localized glycerol-3-phosphate acyltransferase activity protected from all aspects of lipotoxicity. Identification of genes modulating the response to saturated fatty acids may reveal novel therapeutic strategies for treating metabolic diseases linked to lipotoxicity.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Upregulation of SLAMF3 on human T cells is induced by through the STAT5-PI3K/Akt pathway and features the chronic inflammatory profiles of type 2 .

Metabolic stress-induced low-grade chronic inflammation plays an important role in the development of insulin-resistance and type 2 (T2D). Emerging evidence highlights the importance of directly elucidating T-cell activation under the obesity-induced metabolic stress condition, as T cells primed under such conditions were found to acquire a unique phenotype and function. Herein, we found a significant upregulation of signaling lymphocytic activation molecule family member 3 (SLAMF3) expression on T cells from T2D patients compared to those of healthy controls. Importantly, SLAMF3 upregulation was associated with an increased ability to produce proinflammatory cytokines. Significantly increased SLAMF3 expression was seen in T2D patient T cells that produce IFN-γ or IL-17 upon short (4-h) stimulation, compared to non-cytokine-producing T cells. In line with this finding, SLAMF3 T cells were significantly more sensitive than SLAMF3 T cells to TCR stimulation with anti-CD3/CD28 antibodies. Furthermore, treatment with (PA) led to significant upregulation of SLAMF3 on human T cells primed by anti-CD3/CD28 antibodies and on Jurkat cells, a human T-cell line. RNA sequencing revealed strong activation of the PI3K/Akt signaling pathway in T cells that were primed with PA. Further mechanistic studies showed that inhibition of PI3K/Akt signaling, or its upstream mediator STAT5 can prevent PA-induced SLAMF3 upregulation on T cells. These results indicate that SLAMF3 upregulation is associated with T-cell activation and cytokine production in T2D patients, and suggest that elevated saturated fatty acids in T2D patients may induce SLAMF3 upregulation on T cells via activation of the STAT5-PI3K/Akt signaling pathway.

Keyword: diabetes

A Novel Role for Somatostatin in the Survival of Mouse Pancreatic Beta Cells.

Cross-talk between different pancreatic islet cell types regulates islet function and somatostatin (SST) released from pancreatic delta cells inhibits insulin secretion from pancreatic beta cells. In other tissues SST exhibits both protective and pro-apoptotic properties in a tissue-specific manner, but little is known about the impact of the peptide on beta cell survival. Here we investigate the specific role of SST in the regulation of beta cell survival in response to physiologically relevant inducers of cellular stress including palmitate, cytokines and glucose.Pancreatic MIN6 beta cells and primary mouse islet cells were pre-treated with SST with or without the G signalling inhibitor, pertussis toxin, and exposed to different cellular stress factors. Apoptosis and proliferation were assessed by measurement of caspase 3/7 activity, TUNEL and BrdU incorporation, respectively, and expression of target genes was measured by qPCR.SST partly alleviated upregulation of cellular stress markers (Hspa1a and Ddit3) and beta cell apoptosis in response to factors such as lipotoxicity (palmitate), pro-inflammatory cytokines (IL1β and TNFα) and low glucose levels. This effect was mediated via a G protein-dependent pathway, but did not modify transcriptional upregulation of the specific NFκB-dependent genes, Nos2 and Ccl2, nor was it associated with transcriptional changes in SST receptor expression.Our results suggest an underlying protective effect of SST which modulates the beta cell response to ER stress and apoptosis induced by a range of cellular stressors associated with type 2 .© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: diabetes

Mitochondria-targeted therapy rescues development and quality of embryos derived from oocytes matured under oxidative stress conditions: a bovine in vitro model.

Can we use a mitochondrial-targeted antioxidant (Mitoquinone) during in vitro embryo culture to rescue developmental competence of oocytes matured under lipotoxic conditions, exhibiting mitochondrial dysfunction and oxidative stress?Supplementation of embryo culture media with Mitoquinone reduced oxidative stress and prevented mitochondrial uncoupling in embryos derived from metabolically compromised oocytes in vitro, leading to higher blastocyst rates and lower blastomeric apoptosis.Maternal metabolic disorders, such as obesity and type-II are associated with hyperlipidemia and elevated free fatty (FFA) concentrations in the ovarian follicular fluid (FF). Oocyte maturation under these lipotoxic conditions results in increased oxidative stress levels, mitochondrial dysfunction, reduced developmental competence and disappointing IVF results.A well-described bovine oocyte IVM model was used, where a pathophysiologically relevant elevated FF concentrations of (PA; 150\xa0μM or 300\xa0μM) were added to induce oxidative stress. After fertilization (Day 0, D0), zygotes were in vitro cultured (IVC, from D1 to D8) in standard fatty -free media in the presence or absence of Mitoquinone or its carrier triphenyl-phosphonium.Embryo cleavage and fragmentation (D2) and blastocyst rates (D8) were recorded. Mitochondrial activity and oxidative stress in cleaved embryos at D2 were determined using specific fluorogenic probes and confocal microscopy. D8 blastocysts were used to (i) examine the expression of marker genes related to mitochondrial unfolded protein responses (UPRmt; HSPD1 and HSPE1), mitochondrial biogenesis (TFAM), endoplasmic reticulum (ER) UPR (ATF4, ATF6 and BiP) and oxidative stress (CAT, GPX1 and SOD2) using real time RT-PCR; (ii) determine cell differentiation and apoptosis using CDX-2 and cleaved caspase-3 immunostaining; and (iii) measure mtDNA copy numbers. This was tested in a series of experiments with at least three independent replicates for each, using a total of 2525 oocytes. Differences were considered significant if a P value was <0.05 after Bonferroni correction.Exposure to PA during IVM followed by culture under control conditions resulted in a significant increase in oxidative stress in embryos at D2. This was associated with a significant reduction in mitochondrial inner membrane potential (uncoupling) compared with solvent control (P\xa0<\u20090.05). The magnitude of these effects was PA-concentration dependent. Consequently, development to the blastocysts stage was significantly hampered. Surviving blastocysts exhibited high apoptotic cell indices and upregulated mRNA expression indicating persistent oxidative stress, mitochondrial and ER UPRs. In contrast, supplementation of PA-derived zygotes with Mitoquinone during IVC (i) prevented mitochondrial uncoupling and alleviated oxidative stress at D2; and (ii) rescued blastocyst quality; normalized oxidative stress and UPR related genes and apoptotic cell indices (P\xa0>\u20090.01 compared with solvent control). Mitoquinone also improved blastocyst rate in PA-exposed groups, an effect that was dependent on PA concentration.N/A.This is a fundamental study performed using a bovine in vitro model using PA-induced lipotoxicity during oocyte maturation. PA is the most predominant FFA in the FF that is known to induce lipotoxicity; however, in vivo maturation in patients suffering from maternal metabolic disorders involve more factors that cannot be represented in one model. Nevertheless, focusing on the carryover oxidative stress as a known key factor affecting developmental competence, and considering the novel beneficial rescuing effects of Mitoquinone shown here, we believe this model is of high biological relevance.Human oocytes collected for IVF treatments from patients with maternal metabolic disorders are vulnerable to lipotoxicity and oxidative stress during in vivo maturation. The results shown here suggest that mitochondrial targeted therapy, such as using Mitoquinone, during IVC may rescue the developmental competence and quality of these compromised oocytes. After further clinical trials, this may be a valuable approach to increase IVF success rates for infertile patients experiencing metabolic disorders.This study was financially supported by a BOF/KP grant number 34399, from the University of Antwerp, Belgium. W.F.A.M. was supported by a postdoctoral fellowship from the Research Foundation-Flanders (FWO), grant number 12I1417N, Antwerp, Belgium. The Leica SP 8 confocal microscope used in this study was funded by the Hercules Foundation of the Flemish Government (Hercules grant AUHA.15.12). All authors have no financial or non-financial competing interests to declare.© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: diabetes

Glycation of human serum albumin impairs binding to the glucagon-like peptide-1 analogue liraglutide.

The long-acting glucagon-like peptide-1 analogue liraglutide has proven efficiency in the management of type 2 and also has beneficial effects on cardiovascular diseases. Liraglutide\'s protracted action highly depends on its capacity to bind to albumin via its part. However, in , albumin can undergo glycation, resulting in impaired drug binding. Our objective in this study was to assess the impact of human serum albumin (HSA) glycation on liraglutide affinity. Using fluorine labeling of the drug and F NMR, we determined HSA affinity for liraglutide in two glycated albumin models. We either glycated HSA by incubation with glucose (G25- or G100-HSA) or methylglyoxal (MGO-HSA) or purified glycated HSA from the plasma of diabetic patients with poor glycemic control. Nonglycated commercial HSA (G0-HSA) and HSA purified from plasma of healthy individuals served as controls. We found that glycation decreases affinity for liraglutide by 7-fold for G100-HSA and by 5-fold for MGO-HSA compared with G0-HSA. A similarly reduced affinity was observed for HSA purified from diabetic individuals compared with HSA from healthy individuals. Our results reveal that glycation significantly impairs HSA affinity to liraglutide and confirm that glycation contributes to liraglutide\'s variable therapeutic efficiency, depending on stage. Because is a progressive disease, the effect of glycated albumin on liraglutide affinity found here is important to consider when is managed with this drug.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: diabetes

Mangiferin Improved Palmitate-Induced-Insulin Resistance by Promoting Free Fatty Metabolism in HepG2 and C2C12 Cells via PPAR: Mangiferin Improved Insulin Resistance.

Elevated free fatty (FFA) is a key risk factor for insulin resistance (IR). Our previous studies found that mangiferin could decrease serum FFA levels in obese rats induced by a high-fat diet. Our research was to determine the effects and mechanism of mangiferin on improving IR by regulating FFA metabolism in HepG2 and C2C12 cells. The model was used to quantify PA-induced lipid accumulation in the two cell lines treated with various concentrations of mangiferin simultaneously for 24\u2009h. We found that mangiferin significantly increased insulin-stimulated glucose uptake, via phosphorylation of protein kinase B (P-AKT), glucose transporter 2 (GLUT2), and glucose transporter 4 (GLUT4) protein expressions, and markedly decreased glucose content, respectively, in HepG2 and C2C12 cells induced by PA. Mangiferin significantly increased FFA uptake and decreased intracellular FFA and triglyceride (TG) accumulations. The activity of the peroxisome proliferator-activated receptor (PPAR) protein and its downstream proteins involved in fatty translocase (CD36) and carnitine palmitoyltransferase 1 (CPT1) and the fatty -oxidation rate corresponding to FFA metabolism were also markedly increased by mangiferin in HepG2 and C2C12 cells. Furthermore, the effects were reversed by siRNA-mediated knockdown of PPAR. Mangiferin ameliorated IR by increasing the consumption of glucose and promoting the FFA oxidation via the PPAR pathway in HepG2 and C2C12 cells.

Keyword: diabetes

Transport of Ca and Ca-Dependent Permeability Transition in Rat Liver Mitochondria under the Streptozotocin-Induced Type I .

Although is known to be a disease associated with mitochondrial dysfunction, not everything is clear about mitochondrial Ca transport and Ca-induced permeability transition in diabetic cells. The objective of this work was to study the operation of MCU and Ca-dependent mitochondrial permeabilization in the liver cells of Sprague-Dawley rats under the streptozotocin-induced type I . It was shown that two weeks after the induction of , the rate of Ca uptake by the mitochondria of diabetic animals increased ~1.4-fold. The expression of MCU and MICU1 subunits did not change, yet the quantity of dominant-negative MCUb channel subunits was almost twice as lower. The organelles also became more resistant to the induction of CsA-sensitive MPT pore and less resistant to the induction of CsA-insensitive palmitate/Ca-induced pore. The mitochondria of diabetic liver cells also showed changes in the lipid matrix of their membranes. The content of fatty acids in the membranes grew, and microviscosity of the lipid bilayer (assessed with laurdan) increased. At the same time, lipid peroxidation (assessed by the production of malonic dialdehyde) was stimulated. The paper discusses the consequences of the -related changes in mitochondria in the context of cell physiology.

Keyword: diabetes

Taurine improves low-level inorganic arsenic-induced insulin resistance by activating PPARγ-mTORC2 signalling and inhibiting hepatic autophagy.

Inorganic arsenic (iAs) is reportedly associated with the increased incidence of type 2 in the population. Here, we found that iAs exposure significantly decreased the expression of glycolytic genes and glycogen content and increased gluconeogenesis gene levels in C57/BL6J mice. The expression of peroxisome proliferator-activated receptor γ (PPARγ), and mechanistic target of rapamycin complex 2 (mTORC2) were decreased in the livers of iAs-treated mice. Furthermore, in iAs-treated HepG2 cells, we found that PPARγ agonist rosiglitazone (RGS) increased the expression of mTORC2, inhibited autophagy, and improved glucose metabolism. mTORC2 agonist inhibited autophagy and improved glucose metabolism as well as the autophagosome formation inhibitor 3-methyladenine. Taurine, a natural compound, reversed impaired glucose metabolism and decreased expression of PPARγ and mTORC2 induced by iAs in mice liver and HepG2 cells. These data indicated that taurine administration could ameliorate iAs-induced insulin resistance through activating PPARγ-mTORC2 signalling and subsequently inhibiting hepatic autophagy.© 2018 Wiley Periodicals, Inc.

Keyword: diabetes

Triterpenoids Improve -Induced Hepatic Inflammation the Rho-Kinase-Dependent Pathway.

This study aimed to assess the effects of triterpene extract of (Batal.) Iljinskaja (CPT) on -induced hepatic inflammation and to unveil the underlying mechanisms. in db/db mice was alleviated after CPT administration, as assessed by the oral glucose tolerance test. In addition, treatment with CPT dramatically reduced serum insulin, aspartate amino-transaminase, alanine aminotransferase, triglyceride, and total cholesterol amounts. Besides, serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were also reduced after CPT administration. Western blot analysis revealed that CPT treatment significantly reversed the protein expression levels of Rho, ROCK1, ROCK2, p-P65, p-IκBα, p-IKKα, and p-IKKβ in liver samples obtained from db/db mice. Upon stimulation, the protective effects of CPT on the liver were further assessed in HepG2 and LO2 cells, and no appreciable cytotoxic effects were found. Therefore, these findings indicate that CPT alleviates liver inflammation Rho-kinase signaling. Metformin (PubChem CID: 4091); Fasudil (PubChem CID: 3547); (PubChem CID: 985).

Keyword: diabetes

Histone demethylase UTX is a therapeutic target for diabetic kidney disease.

Diabetic kidney disease (DKD) is a major complication of . We found that UTX (ubiquitously transcribed tetratricopeptide repeat on chromosome X, also known as KDM6A), a histone demethylase, was upregulated in the renal mesangial and tubular cells of diabetic mice and DKD patients. In cultured renal mesangial and tubular cells, UTX overexpression promoted -induced elevation of inflammation and DNA damage, whereas UTX knockdown or GSK-J4 treatment showed the opposite effects. We found that UTX demethylase activity-dependently regulated the transcription of inflammatory genes and apoptosis; moreover, UTX bound with p53 and p53-dependently exacerbated DNA damage. Administration of GSK-J4, an H3K27 demethylase inhibitor, ameliorated the -induced renal abnormalities in db/db mice, an animal model of type 2 . These results revealed the possible mechanisms underlying the regulation of histone methylation in DKD and suggest UTX as a potential therapeutic target for DKD.Diabetic kidney disease (DKD) is a microvascular complication of and the leading cause of end-stage kidney disease worldwide without effective therapy available. UTX (ubiquitously transcribed tetratricopeptide repeat on chromosome X, also known as KDM6A), a histone demethylase that removes the di- and tri-methyl groups from histone H3K27, plays important biological roles in gene activation, cell fate control and life span regulation in Caenorhabditis elegans. In the present study, we report upregulated UTX in the kidneys of diabetic mice and DKD patients. Administration of GSK-J4, an H3K27 demethylase inhibitor, ameliorated the -induced renal dysfunction, abnormal morphology, inflammation, apoptosis and DNA damage in db/db mice, comprising an animal model of type 2 . In cultured renal mesanglial and tubular cells, UTX overexpression promoted induced elevation of inflammation and DNA damage, whereas UTX knockdown or GSK-J4 treatment showed the opposite effects. Mechanistically, we found that UTX demethylase activity-dependently regulated the transcription of inflammatory genes; moreover, UTX bound with p53 and p53-dependently exacerbated DNA damage. Collectively, our results suggest UTX as a potential therapeutic target for DKD.© 2018 The Authors. The Journal of Physiology © 2018 The Physiological Society.

Keyword: diabetes

Phytochemical Profiling, GC-MS Analysis and α-Amylase Inhibitory Potential of Ethanolic Extract of Linn. Endocarp.

Drugs with post-prandial action constitute one of the main courses of treatments for .In the present investigation, we have explored the α-amylase inhibitory potential of ethanolic extract of Cocos nucifera endocarp.DNS based assay was done to assess the α-amylase inhibition potential of ethanolic extract. Phytochemical screening and GC-MS analysis were done in order to assess the chemical profiling of extract. In silico docking studies were done using VLife MDS 4.6 software and the probable molecules, predicted after GC-MS analysis, were docked with the co-crystallized (acarbose) tracked active site and rest all cavities of porcine pancreatic α-amylase (1OSE). ADMET analysis was done using StarDrop 6.4, Derek Nexus and P450 Modules from Optibrium Ltd. and LHASA Ltd.DNS based α-amylase assay indicated that the IC50 value of extract lies in the range of 63- 126 µg/ml and at higher doses, i.e. above 250 µg/ml, it has better α-amylase inhibition than the standard drug, acarbose. Phytochemical screening indicated that ethanolic extract is rich in alkaloids, tannins, flavonoids, saponins, triterpenes, glycosides, carbohydrates, terpenoids, quinones and lactones. Further, GC-MS analysis (where Similarity Index was > 90) predicted that the probable phytoconstituents present in the ethanolic extract are myristic , syringaldehyde, eugenol, vanillin, 2,4-di-tert-butylphenol, lauric , methyl ester and γ-sitosterol. γ-Sitosterol showed the strong affinity towards the active site which was tracked by a co-crystallized ligand along with cavity 1 and 2 while significant interactions were observed in case of co-crystallized tracked active site as well as cavity 4 of 1OSE. Ethanolic extract of C. nucifera has no hemolytic effect.Its ability to effectively inhibit α-amylase may be attributed to the presence of the above probable molecules, which will be explored further.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: diabetes

Vascular endothelial growth factor over-expressed mesenchymal stem cells-conditioned media ameliorate palmitate-induced diabetic endothelial dysfunction through PI-3K/AKT/m-TOR/eNOS and p38/MAPK signaling pathway.

In the pathogenesis of (DM), islet microvasculares are severely damaged due to glucolipotoxicity and other reasons. Vascular endothelial growth factor (VEGF) is an indispensable and specific angiogenic factor in the pathogenesis and treatment of diabetic islet microvascular disease. Mesenchymal stem cells (MSCs) are regarded as a promising treatment of because of their immunosuppressive effect and multipotential differentiation potency. In this study, we tested whether MSCs over-expressing VEGF conditioned medium (MSC-VEGF-CM) could ameliorate pancreatic islet endothelial cells (MS-1) dysfunction induced by a common diabetic inducer palmitate (PA). We found that cell survival and migration were restrained by PA and partly repaired by the pro-protected of MSC-VEGF-CM. Meanwhile, PI-3K/AKT/m-TOR/eNOS and p38/MAPK signaling pathways were also up-regulated. Though apoptosis-related proteins, caspase-3 and caspase-9, had no significantly suppressed between MSC-VEGF-CM and MSC-CM alone, the expression levels of vascular surface factors such as CD31, VE-cadherin, occludin and ICAM-1, were remarkably up-regulated by the pro-protected of MSC-VEGF-CM. Our data suggested that MSC-VEGF-CM had therapeutic effect on the PA-induced dysfunction through the re-activation of PI-3K/AKT/m-TOR/eNOS and p38/MAPK signaling pathways.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Systematic review of palm oil consumption and the risk of cardiovascular disease.

The high amount of saturated fatty acids (SFA) coupled with the rising availability and consumption of palm oil have lead to the assumption that palm oil contributes to the increased prevalence of cardiovascular diseases worldwide. We aimed at systematically synthesising the association of palm oil consumption with cardiovascular disease risk and cardiovascular disease-specific mortality.We systematically searched Central, Medline and Embase databases up to June 2017 without restriction on setting or language. We performed separate searches based on the outcomes: coronary heart disease and stroke, using keywords related to these outcomes and palm oil. We searched for published interventional and observational studies in adults (Age: >18 years old). Two investigators extracted data and a consensus was reached with involvement of a third. Only narrative synthesis was performed for all of the studies, as the data could not be pooled.Our search retrieved 2,738 citations for stroke with one included study and 1,777 citations for coronary heart disease (CHD) with four included studies. was reported to be associated with risk of myocardial infarction (MI) (OR 2.76; 95%CI = 1.39-5.47). Total SFA intake was reported to be not significant for risk of MI. Varying intake of fried foods, highest contributor to total SFA with 36% of households using palm oil for frying, showed no significant associations to risk of MI. Odds of developing first non-fatal acute MI was higher in palm oil compared to soybean oil with 5% trans-fat (OR = 1.33; 95%CI = 1.09-1.62) than palm oil compared to soybean oil with 22% trans-fat (OR = 1.16; 95%CI = 0.86-1.56). Nevertheless, these risk estimates were non-significant and imprecise. The trend amongst those taking staple pattern diet (characterised by higher palm oil, red meat and added sugar consumption) was inconsistent across the factor score quintiles. During the years of 1980 and 1997, for every additional kilogram of palm oil consumed per-capita annually, CHD mortality risk was 68 deaths per 100,000 (95% CI = 21-115) in developing countries and 17 deaths per 100,000 (95%CI = 5.3-29) in high-income countries, whereas stroke was associated with 19 deaths per 100,000 (95%CI = -12-49) and 5.1 deaths per 100,000 (95% CI: -1.2-11) respectively. The evidence for the outcomes of this review were all graded as very low. The findings of this review should be interpreted with some caution, owing to the lack of a pooled effect estimate of the association, significant bias in selection criteria and confounding factors, inclusion of other food items together with palm oil, and the possible out-dated trend in the ecological study.In view of the abundance of palm oil in the market, quantifying its true association with CVD outcomes is challenging. The present review could not establish strong evidence for or against palm oil consumption relating to cardiovascular disease risk and cardiovascular disease-specific mortality. Further studies are needed to establish the association of palm oil with CVD. A healthy overall diet should still be prioritised for good cardiometabolic health.

Keyword: diabetes

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced . However, it is unknown if 5-PAHSA affects glucose and lipid metabolism in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid metabolism in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid metabolism. We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid metabolism and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: diabetes

Human embryonic stem cell-derived cardiomyocytes as an in vitro model to study cardiac insulin resistance.

Patients with type 2 (T2D) and/or insulin resistance (IR) have an increased risk for the development of heart failure (HF). Evidence indicates that this increased risk is linked to an altered cardiac substrate preference of the insulin resistant heart, which shifts from a balanced utilization of glucose and long-chain fatty acids (FAs) towards an almost complete reliance on FAs as main fuel source. This shift leads to a loss of endosomal proton pump activity and increased cardiac fat accumulation, which eventually triggers cardiac dysfunction. In this review, we describe the advantages and disadvantages of currently used in vitro models to study the underlying mechanism of IR-induced HF and provide insight into a human in vitro model: human embryonic stem cell-derived cardiomyocytes (hESC-CMs). Using functional metabolic assays we demonstrate that, similar to rodent studies, hESC-CMs subjected to 16h of high palmitate (HP) treatment develop the main features of IR, i.e., decreased insulin-stimulated glucose and FA uptake, as well as loss of endosomal acidification and insulin signaling. Taken together, these data propose that HP-treated hESC-CMs are a promising in vitro model of lipid overload-induced IR for further research into the underlying mechanism of cardiac IR and for identifying new pharmacological agents and therapeutic strategies. This article is part of a Special issue entitled Cardiac adaptations to obesity, and insulin resistance, edited by Professors Jan F.C. Glatz, Jason R.B. Dyck and Christine Des Rosiers.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: diabetes

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 . However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut microbiota and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: diabetes

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 fat cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on lipid-induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and lipid metabolism in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose metabolism, such as insulin receptor () and insulin receptor substrate 1 and 2 ( and ), as well as those involved in lipid metabolism, including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Keyword: diabetes

Identifying Signalling Pathways Regulated by GPRC5B in β-Cells by CRISPR-Cas9-Mediated Genome Editing.

CRISPR-Cas9, a RNA-guided targeted genome editing tool, has revolutionized genetic engineering by offering the ability to precisely modify DNA. GPRC5B is an orphan receptor belonging to the group C family of G protein-coupled receptors (GPCRs). In this study, we analysed the functional roles of the Gprc5b receptor in MIN6 β-cells using CRISPR-Cas9 and transient over-expression of Gprc5b.The optimal transfection reagent for use in MIN6 β-cells was determined by analysing efficiency of GFP plasmid delivery by cell sorting. A MIN6 β-cell line in which Gprc5b expression was knocked down (Gprc5b KD) was generated using CRISPR-Cas9 technology. Gprc5b receptor mRNA expression, proliferation, apoptosis, Cignal 45-Pathway Reporter Array signalling and western blot assays were carried out using Gpcr5b KD MIN6 β-cells that had been transiently transfected with different concentrations of mouse Gprc5b plasmid to over-express Gprc5b.JetPRIME® was the best candidate for MIN6 β-cell transfection, providing approximately 30% transfection efficiency. CRISPR-Cas9 technology targeting Gprc5b led to stable knock-down of this receptor in MIN6 β-cells and its re-expression induced proliferation and potentiated cytokine- and palmitate-induced apoptosis. The Cignal 45 Reporter analysis indicated Gprc5b-dependent regulation of apoptotic and proliferative pathways, and western blotting confirmed activation of signalling via TGF-β and IFNγ.This study provides evidence of CRISPR-Cas9 technology being used to down-regulate Gprc5b expression in MIN6 β-cells. This strategy allowed us to identify signalling pathways linking GPRC5B receptor expression to β-cell proliferation and apoptosis.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Telmisartan protects against high glucose/high lipid-induced apoptosis and insulin secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high lipid (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of . In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of . In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high lipid-induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: diabetes

CTRP3 attenuates high-fat diet-induced male reproductive dysfunction in mice.

Recent studies have suggested a role for abdominal obesity in male infertility. Previous studies have found that cell apoptosis exerts an important role in obesity-related male infertility. C1q/TNF-related protein 3 (CTRP3), a paralog of adiponectin, has been proposed to exert anti-apoptotic effects and to attenuate -related cardiac injuries. However, the role of CTRP3 in high-fat diet (HFD)-induced spermatogenic impairment remains unclear. In the present study, we fed male mice an HFD for 24 weeks to induce obesity. The expression of CTRP3 was decreased by HFD feeding. Supplementation with the recombinant human globular domain of CTRP3 (0.25 μg/g/day) for 4 weeks beginning at 20 weeks of the HFD improved spermatogenic function in the HFD-fed mice, which were characterized by improved testis morphology, increased testis weight/body weight ratio, and increased sperm count, sperm viability, and sperm motility. We also found that CTRP3 infusion resulted in the attenuation of endoplasmic reticulum (ER) stress and the activation of silence information regulator 1 (SIRT1) in the testes of obese mice. Our study also suggested that CTRP3 attenuated the (PA)-induced reductions in sperm viability and motility via the inhibition of ER stress. Moreover, germ cell-specific knockout abolished the protective effects of CTRP3 and studies of human sperm showed that the protective effects of CTRP3 on sperm viability and motility were abrogated by a specific inhibitor of SIRT1. Thus, our results demonstrated that CTRP3 expression protected against HFD-induced spermatogenic deficiency through the SIRT1/ER stress pathway.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: diabetes

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the metabolism of fatty acids (FA) and only in the second turn in glucose metabolism; regulation of FA metabolism in cells started millions of years earlier than that of glucose metabolism. Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA metabolism instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I , 2. rare structurally-related (receptor pathology) hyperinsulinemic type II , and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, metabolic syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: diabetes

Microfluidic-enabled quantitative measurements of insulin release dynamics from single islets of Langerhans in response to 5- hydroxy stearic .

Proper release of insulin from pancreatic islets of Langerhans is essential for maintaining glucose homeostasis. For full efficacy, both the pattern and the amount of hormone release are critical. It is therefore important to understand how insulin levels are secreted from single islets in both a quantitative fashion and in a manner that resolves temporal dynamics. In this study, we describe a microfluidic analytical system that can both quantitatively monitor insulin secretion from single islets while simultaneously maintaining high temporal sampling to resolve dynamics of release. We have applied this system to determine the acute and chronic effects of a recently-identified lipid, 5- hydroxy stearic (5-PAHSA), which is a member of the fatty hydroxy fatty class of lipids that are upregulated in healthy individuals. Chronic incubation (48 h) with 5-PAHSA significantly increased glucose-stimulated insulin secretion (GSIS) in murine islets compared to chronic incubation without the lipid or in the presence of (PA). The studies were continued in human islets from both healthy donors and donors diagnosed with type 2 (T2DM). Total amounts of GSIS were not only augmented in islets that were chronically incubated with 5-PAHSA, but the dynamic insulin release profiles also improved as noted by more pronounced insulin oscillations. With this quantitative microfluidic system, we have corroborated the anti-diabetic effects of 5-PAHSA by demonstrating improved islet function after chronic incubation with this lipid via improved oscillatory dynamics along with higher basal and peak release rates.

Keyword: diabetes

Antioxidant and antidiabetic activity of blackberry after gastrointestinal digestion and human gut microbiota fermentation.

Blackberry fruit contains high levels of polyphenols particularly anthocyanins which contribute to its biological activities. Bioavailability of polyphenols especially anthocyanins is generally low, it has been proposed that metabolites from polyphenol biotransformation under colonic fermentation are components that exert health benefits. In this study, blackberry was subjected to simulated gastrointestinal digestion and gut microbiota fermentation at different time intervals (0-48\u202fh) to study the changes in bioactive components, its antioxidant and antidiabetic activities. Phenolic compounds, during digestion and fermentation were also analysed. Gut metabolites of blackberry significantly increased the glucose consumption and glycogen content in HepG2 cells. Furthermore, gut metabolites ameliorated high glucose plus -induced ROS overproduction, mitochondrial membrane collapse, and glutathione depletion in HepG2 cells. The mechanism of antidiabetic activity of blackberry was via its potent antioxidant activity. Therefore, our results suggest that blackberry could be recommended as a functional food due to potential antioxidant and antidiabetic activity.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Islet protection and amelioration of type 2 by treatment with quercetin from the flowers of .

The traditional Chinese medicine - the flower of - is reported as an effective therapeutic for type 2 (T2DM). Nevertheless, most constituents of the flowers of have not yet been studied. This study was conducted to investigate the effect of quercetin extracted from the flowers of on islet protection and amelioration in T2DM and explore its mechanism.Quercetin was extracted from the flowers of and verified by high-performance liquid chromatography. Quercetin or crude extract\'s effect on insulin secretion was investigated. ERK1/2 and phospho-ERK1/2 were detected by Western blot analysis, and fluo-3 AM was used to detect intracellular Ca. The anti-apoptosis effect of quercetin or crude extract on MIN-6 cells was investigated by thiazolyl blue tetrazolium bromide (MTT) assay and flow cytometry analysis. Activation of caspases and expression of Bcl-2 and BAX were tested by Western blot analysis. In addition, the mitochondrial membrane potential was determined by JC-1 probe. Moreover, in vivo activity was also tested in db/db mice.A quercetin level of >10 μmol/L could induce insulin secretion. Intracellular Ca and ERK1/2 were involved in the signaling pathway of quercetin-induced insulin secretion. We also observed that quercetin could inhibit -induced cell apoptosis via suppressing the activation of caspase-3, -9, -12; increasing the ratio of Bcl-2/BAX and reversing the impaired mitochondrial membrane potential. Crude extract\'s effect on insulin secretion was similar to that of pure extracted quercetin, while it possessed higher anti-apoptosis activity. Additionally, intraperitoneal glucose tolerance, plasma insulin level, hepatic triglyceride, hepatic glycogen and the pathological histology of both pancreatic islet and liver in db/db mice were significantly improved by the administration of the extracted quercetin.Our study indicated that quercetin extracted from the flowers of exerted excellent properties in islet protection and amelioration.

Keyword: diabetes

Downregulation of MALAT1 alleviates saturated fatty -induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis.

The increased level of saturated fatty acids (SFAs) is found in patients with , obesity, and other metabolic disorders. SFAs can induce lipotoxic damage to cardiomyocytes, but the mechanism is unclear. The long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acts as a key regulator in (PA)-induced hepatic steatosis, but its role in PA-induced myocardial lipotoxic injury is still unknown. The aim of this study was to explore the role and underlying mechanism of MALAT1 in PA-induced myocardial lipotoxic injury. MALAT1 expression in PA-treated human cardiomyocytes (AC16 cells) was detected by RT-qPCR. The effect of MALAT1 on PA-induced myocardial injury was measured by Cell Counting Kit-8, lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB) assays. Apoptosis was detected by flow cytometry. The activities of cytokines and nuclear factor (NF)-κB were detected by enzyme-linked immunosorbent assay. The interaction between MALAT1 and miR-26a was evaluated by a luciferase reporter assay and RT-qPCR. The regulatory effects of MALAT1 on high mobility group box 1 (HMGB1) expression were evaluated by RT-qPCR and western blotting. MALAT1 was significantly upregulated in cardiomyocytes after PA treatment. Knockdown of MALAT1 increased the viability of PA-treated cardiomyocytes, decreased apoptosis, and reduced the levels of LDH, CK-MB, TNF-α, and IL-1β. Moreover, we found that MALAT1 specifically binds to miR-26a and observed a reciprocal negative regulatory relationship between these factors. We further found that the downregulation of MALAT1 represses HMGB1 expression, thereby inhibiting the activation of the Toll-like receptor 4 (TLR4)/NF-κB-mediated inflammatory response. These repressive effects were rescued by an miR-26a inhibitor. We demonstrate that MALAT1 is induced by SFAs and its downregulation alleviates SFA-induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis. Our findings provide new insight into the mechanism underlying myocardial lipotoxic injury.

Keyword: diabetes

High-density lipoprotein ameliorates -induced lipotoxicity and oxidative dysfunction in H9c2 cardiomyoblast cells via ROS suppression.

High levels circulating saturated fatty acids are associated with , obesity and hyperlipidemia. In heart, the accumulation of saturated fatty acids has been determined to play a role in the development of heart failure and diabetic cardiomyopathy. High-density lipoprotein (HDL) has been reported to possess key atheroprotective biological properties, including cellular cholesterol efflux capacity, anti-oxidative and anti-inflammatory activities. However, the underlying mechanisms are still largely unknown. Therefore, the aim of the present study is to test whether HDL could protect (PA)-induced cardiomyocyte injury and explore the possible mechanisms.H9c2 cells were pretreated with HDL (50-100\u2009μg/ml) for 2\u2009h followed by PA (0.5\u2009mM) for indicated time period. Our results showed that HDL inhibited PA-induced cell death in a dose-dependent manner. Moreover, HDL rescued PA-induced ROS generation and the phosphorylation of JNK which in turn activated NF-κB-mediated inflammatory proteins expressions. We also found that PA impaired the balance of BCL family proteins, destabilized mitochondrial membrane potential, and triggered subsequent cytochrome c release into the cytosol and activation of caspase 3. These detrimental effects were ameliorated by HDL treatment.PA-induced ROS accumulation and results in cardiomyocyte apoptosis and inflammation. However, HDL attenuated PA-induced lipotoxicity and oxidative dysfunction via ROS suppression. These results may provide insight into a possible molecular mechanism underlying HDL suppression of the free fatty -induced cardiomyocyte apoptosis.

Keyword: diabetes

Fibroblast growth factor 21 protects against lipotoxicity-induced pancreatic β-cell dysfunction via regulation of AMPK signaling and lipid metabolism.

Fibroblast growth factor 21 (FGF21) is known as a potent metabolic regulator but its protective mechanisms against lipotoxicity-induced β-cell dysfunction and apoptosis remain elusive. Here, we aimed to examine the regulatory pathways whereby FGF21 mediates islet lipid metabolism in lipotoxicity-treated cells and animal models. Rat β-cell line (INS-1E cells) and islets isolated from C57/BL6J mice were exposed to (PA) with/without FGF21, mimicking lipotoxic conditions. Resultant insulin secretion and intracellular signaling were analyzed with Western blotting and RNA-seq. C57/BL6J and global FGF21 knockout (KO) mice were fed with a high-fat diet (HFD) to induce lipotoxicity and given with a long-acting mimetic of FGF21. Insulin resistance and β-cell function were then assessed using homeostasis model assessment of insulin resistance (HOMA-IR) and insulinogenic index. FGF21 ameliorated PA-induced lipid accumulation, reversed cell apoptosis, and enhanced glucose-stimulated insulin secretion (GSIS) as impaired by lipotoxicity in islet β-cells. Mechanistically, FGF21 exerted its beneficial effects through activation of AMPK-ACC (acetyl-CoA carboxylase) pathway and peroxisome proliferation-activated receptors (PPARs) δ/γ signaling, thus increasing the levels of carnitine palmitoyltransferase-1A (CPT1A) and leading to increased fatty (FA) oxidation and reduced lipid deposition in β-cells. Interestingly, FGF21 reduced PA-induced cell death via restoration of the expression of apoptosis inhibitor Birc3. In vivo studies further showed that FGF21 is critical for islet insulinogenic capacity and normal function in the context of HFD-treated animals. FGF21 down-regulates islet cell lipid accumulation, probably via activation of AMPK-ACC and PPARδ/γ signaling, and reduces cell death under lipotoxicity, indicating that FGF21 is protective against lipotoxicity-induced β-cell dysfunction and apoptosis.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: diabetes

Role of the saturated fatty palmitate in the interconnected hypothalamic control of energy homeostasis and biological rhythms.

The brain, specifically the hypothalamus, controls whole body energy and glucose homeostasis through neurons that synthesize specific neuropeptides, whereas hypothalamic dysfunction is linked directly to insulin resistance, obesity, and type 2 . Nutrient excess, through overconsumption of a Western or high-fat diet, exposes the hypothalamus to high levels of free fatty acids, which induces neuroinflammation, endoplasmic reticulum stress, and dysregulation of neuropeptide synthesis. Furthermore, exposure to a high-fat diet also disrupts normal circadian rhythms, and conversely, clock gene knockout models have symptoms of metabolic disorders. While whole brain/animal studies have provided phenotypic end points and important clues to the genes involved, there are still major gaps in our understanding of the intracellular pathways and neuron-specific components that ultimately control circadian rhythms and energy homeostasis. Because of its complexity and heterogeneous nature, containing a diverse mix cell types, it is difficult to dissect the critical hypothalamic components involved in these processes. Of significance, we have the capacity to study these individual components using an extensive collection of both embryonic- and adult-derived, immortalized hypothalamic neuronal cell lines from rodents. These defined neuronal cell lines have been used to examine the impact of nutrient excess, such as palmitate, on circadian rhythms and neuroendocrine signaling pathways, as well as changes in vital neuropeptides, leading to the development of neuronal inflammation; the role of proinflammatory molecules in this process; and ultimately, restoration of normal signaling, clock gene expression, and neuropeptide synthesis in disrupted states by beneficial anti-inflammatory compounds in defined hypothalamic neurons.

Keyword: diabetes

Neuroprotective effects of vitamin D on high fat diet- and -induced enteric neuronal loss in mice.

The role of vitamin D in obesity and is debated. Obese and/or diabetic patients have elevated levels of free fatty acids, increased susceptibility to gastrointestinal symptoms and are suggested to have altered vitamin D balance. The enteric nervous system is pivotal in regulating gastrointestinal activity and high fat diet (HFD) has been shown to cause loss of enteric neurons in ileum and colon. This study investigates the effect of vitamin D on HFD- and -induced enteric neuronal loss in vivo and in vitro.Mice were fed either a normal diet (ND) or HFD supplemented with varying levels of vitamin D (from 0x to 20x normal vitamin D level) for 19 weeks. Ileum and colon were analyzed for neuronal numbers and remodeling. Primary cultures of myenteric neurons from mouse small intestine were treated with (4x10M) and/or 1α,25-hydroxy-vitamin D3 (VD, 10- 10M) with or without modulators of lipid metabolism and VD pathways. Cultures were analyzed by immunocyto- and histochemical methods.Vitamin D supplementation had no effect on enteric neuronal survival in the ND group. HFD caused substantial loss of myenteric neurons in ileum and colon. Vitamin D supplementation between 0-2x normal had no effect on HFD-induced neuronal loss. Supplementation with 20x normal, prevented the HFD-induced neuronal loss. In vitro supplementation of VD prevented the -induced neuronal loss. The VD receptor (VDR) was not identified in enteric neurons. Enteric glia expressed the alternative VD receptor, protein disulphide isomerase family A member 3 (PDIA3), but PDIA3 was not found to mediate the VD response in vitro. Inhibition of peroxisome proliferator-activated receptor gamma (PPARγ) and immune neutralization of isocitrate lyase prevented the VD mediated neuroprotection to exposure.Results show that VD protect enteric neurons against HFD and induced neuronal loss. The mechanism behind is suggested to be through activation of PPARγ leading to improved neuronal peroxisome function and metabolism of neuronal lipid intermediates.

Keyword: diabetes

Protective potential of klotho protein on diabetic retinopathy: Evidence from clinical and in\xa0vitro studies.

The purpose of the present study was to observe the relationship between serum α-klotho (KL) protein level and diabetic retinopathy (DR), and to further examine the effects of KL protein on apoptosis induced by (PA) in human retinal endothelial cells.A total of 17 healthy people and 60 type\xa02 patients were included. According to the results from fundus fluorescein angiography, the patients were divided into three subgroups: without DR, non-proliferative DR and proliferative DR. Serum KL level was measured by enzyme-linked immunosorbent assay. In\xa0vitro, human retinal endothelial cells were exposed to PA with or without KL protein. Apoptosis rates were analyzed by flow cytometry analysis. Apoptotic-related protein expressions were detected by western blotting analysis.Serum KL level was lower in patients than that in healthy participants (P\xa0=\xa00.007), and was gradually decreased among the without DR, non-proliferative DR and proliferative DR subgroups (P\xa0=\xa00.045). A logistic regression analysis showed that after adjusting for the other confounding factors, serum KL level was independently and negatively related with DR (P\xa0=\xa00.049). Furthermore, the increased apoptosis rates induced by PA were inhibited with the addition of KL protein. Consistently, KL protein reversed the expression levels of the increased pro-apoptotic protein Bax and the decreased anti-apoptotic protein Bcl-2 induced by PA. However, the anti-apoptotic effect of KL protein was attenuated by LY294002 through the phosphatidylinositol 3 kinase-serine∕threonine kinase pathway.The data suggested that KL protein was probably a potential protective factor against retinopathy in type\xa02 patients.© 2019 The Authors. Journal of Investigation published by Asian Association for the Study of (AASD) and John Wiley & Sons Australia, Ltd.

Keyword: diabetes

High Fat Diet Upregulates Fatty Oxidation and Ketogenesis via Intervention of PPAR-γ.

Systemic hyperlipidemia and intracellular lipid accumulation induced by chronic high fat diet (HFD) leads to enhanced fatty oxidation (FAO) and ketogenesis. The present study was aimed to determine whether activation of peroxisome proliferator-activated receptor-γ (PPAR-γ) by surplus free fatty acids (FA) in hyperlipidemic condition, has a positive feedback regulation over FAO and ketogenic enzymes controlling lipotoxicity and cardiac apoptosis.8 weeks old C57BL/6 wild type (WT) or PPAR-γ-/- mice were challenged with 16 weeks 60% HFD to induce obesity mediated type 2 (T2DM) and diabetic cardiomyopathy. Treatment course was followed by echocardiographic measurements, glycemic and lipid profiling, immunoblot, qPCR and immunohistochemistry (IHC) analysis of PPAR-γ and following mitochondrial metabolic enzymes 3-hydroxy-3-methylglutaryl-CoA synthase (HMGCS2), mitochondrial β- hydroxy butyrate dehydrogenase (BDH1) and pyruvate dehydrogenase kinase isoform 4 (PDK4). In vivo model was translated in vitro, with neonatal rat cardiomyocytes (NRCM) treated with PPAR-γ agonist/antagonist and PPAR-γ overexpression adenovirus in presence of (PA). Apoptosis was determined in vivo from left ventricular heart by TUNEL assay and immunoblot analysis.We found exaggerated circulating ketone bodies production and expressions of the related mitochondrial enzymes HMGCS2, BDH1 and PDK4 in HFD-induced diabetic hearts and in PA-treated NRCM. As a mechanistic approach we found HFD mediated activation of PPAR-γ is associated with the above-mentioned mitochondrial enzymes. HFD-fed PPAR-γ-/-mice display decreased hyperglycemia, hyperlipidemia associated with increased insulin responsiveness as compared to HFD-fed WT mice PPAR-γ-/-HFD mice demonstrated a more robust functional recovery after induction, as well as significantly reduced myocyte apoptosis and improved cardiac function.PPAR-γ has been described previously to regulate lipid metabolism and adipogenesis. The present study suggests for the first time that increased PPAR-γ expression by HFD is responsible for cardiac dysfunction via upregulation of mitochondrial enzymes HMGCS2, BDH1 and PDK4. Targeting PPAR-γ and its downstream mitochondrial enzymes will provide novel strategies in preventing metabolic and myocardial dysfunction in .© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Lupin seed hydrolysate promotes G-protein-coupled receptor, intracellular Ca and enhanced glycolytic metabolism-mediated insulin secretion from BRIN-BD11 pancreatic beta cells.

Lupin seed proteins have been reported to exhibit hypoglycaemic effects in animals and humans following oral administration, however little is known about its mechanism of action. This study investigated the signalling pathway(s) responsible for the insulinotropic effect of the hydrolysate obtained from lupin (Lupinus angustifolius L.) seed extracts utilizing BRIN-BD11 β-cells. The extract was treated with digestive enzymes to give a hydrolysate rich in biomolecules ≤7\u202fkDa. Cells exhibited hydrolysate induced dose-dependent stimulation of insulin secretion and enhanced intracellular Ca and glucose metabolism. The stimulatory effect of the hydrolysate was potentiated by depolarizing concentrations of KCl and was blocked by inhibitors of the ATP sensitive K channel, Gα protein, phospholipase C (PLC) and protein kinase C (PKC). These findings reveal a novel mechanism for lupin hydrolysate stimulated insulin secretion via Gα mediated signal transduction (Gα/PLC/PKC) in the β-cells. Thus, lupin hydrolysates may have potential for nutraceutical treatment in type 2 .Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Metabolic flexibility to lipid availability during exercise is enhanced in individuals with high insulin sensitivity.

Metabolic flexibility to lipid (MetFlex-lip) is the capacity to adapt lipid oxidation to lipid availability. Hypothetically, impaired MetFlex-lip in skeletal muscle induces accumulation of lipid metabolites that interfere with insulin signaling. Our aim was to compare MetFlex-lip during exercise in subjects with low (Low_IS) vs. high (High_IS) insulin sensitivity. Twenty healthy men were designated as Low_IS or High_IS on the basis of the median of the homeostatic model assessment of insulin resistance index. Groups had similar age, body mass index, and maximum oxygen uptake (V̇o). Subjects cycled at 50% V̇o until expending 650 kcal. Adaptation in lipid oxidation was calculated as the drop in respiratory quotient (RQ) at the end of exercise vs. the maximum RQ (ΔRQ). Lipid availability was calculated as the increase in circulating nonesterified fatty acids (NEFA) at the end of exercise vs. the minimum NEFA (ΔNEFA). ΔRQ as a function of ΔNEFA was used to determine MetFlex-lip. On average, RQ and circulating NEFA changed similarly in both groups. However, ΔRQ correlated with ΔNEFA in High_IS ( r\u2009=\u2009-0.83, P < 0.01) but not in Low_IS ( r\u2009=\u2009-0.25, P = 0.48) subjects. Thus the slope of the ΔRQ vs. ΔNEFA relationship was steeper in High_IS vs. Low_IS subjects (-0.139 ± 0.03 vs. -0.025 ± 0.03 RQ·mmol·l, respectively; P < 0.05), with similar intercepts. We conclude that in subjects with High_IS lipid-to-carbohydrate oxidation ratio adapts to the increased circulating NEFA availability during exercise. Such MetFlex-lip appears impaired in subjects with Low_IS. Whether a cause-effect relationship exists between impaired MetFlex-lip and low insulin sensitivity remains to be determined.

Keyword: diabetes

GPR120 protects lipotoxicity-induced pancreatic β-cell dysfunction through regulation of PDX1 expression and inhibition of islet inflammation.

G-protein coupled receptor 120 (GPR120) has been shown to act as an omega-3 unsaturated fatty sensor and is involved in insulin secretion. However, the underlying mechanism in pancreatic β cells remains unclear. To explore the potential link between GPR120 and β-cell function, its agonists docosahexaenoic (DHA) and GSK137647A were used in (PA)-induced pancreatic β-cell dysfunction, coupled with GPR120 knockdown (KD) in MIN6 cells and GPR120 knockout (KO) mice to identify the underlying signaling pathways. and treatments of MIN6 cells and islets isolated from wild-type (WT) mice with DHA and GSK137647A restored pancreatic duodenal homeobox-1 (PDX1) expression levels and β-cell function via inhibiting PA-induced elevation of proinflammatory chemokines and activation of nuclear factor κB, c-Jun amino (N)-terminal kinases1/2 and p38MAPK signaling pathways. On the contrary, these GPR120 agonism-mediated protective effects were abolished in GPR120 KD cells and islets isolated from GPR120 KO mice. Furthermore, GPR120 KO mice displayed glucose intolerance and insulin resistance relative to WT littermates, and β-cell functional related genes were decreased while inflammation was exacerbated in islets with increased macrophages in pancreas from GPR120 KO mice. DHA and GSK137647A supplementation ameliorated glucose tolerance and insulin sensitivity, as well as improved expression and islet inflammation in diet-induced obese WT mice, but not in GPR120 KO mice. These findings indicate that GPR120 activation is protective against lipotoxicity-induced pancreatic β-cell dysfunction, via the mediation of PDX1 expression and inhibition of islet inflammation, and that GPR120 activation may serve as a preventative and therapeutic target for obesity and .© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: diabetes

Saturated fatty stimulates production of extracellular vesicles by renal tubular epithelial cells.

Lipotoxicity, an accumulation of intracellular lipid metabolites, has been proposed as an important pathogenic mechanism contributing to kidney dysfunction in the context of metabolic disease. , a predominant lipid derivative, can cause lipoapoptosis and the release of inflammatory extracellular vesicles (EVs) in hepatocytes, but the effect of lipids on EV production in chronic kidney disease remains vaguely explored. This study was aimed to investigate whether would stimulate EV release from renal proximal tubular epithelial cells. Human and rat proximal tubular epithelial cells, HK-2 and NRK-52E, were incubated with 1% bovine serum albumin (BSA), BSA-conjugated (PA), and BSA-conjugated oleic (OA) for 24-48\xa0h. The EVs released into conditioned media were isolated by ultracentrifugation and quantified by nanoparticle-tracking analysis (NTA). According to NTA, the size distribution of EVs was 30-150\xa0nm with similar mode sizes in all experimental groups. Moreover, BSA-induced EV release was significantly enhanced in the presence of PA, whereas EV release was not altered by the addition of OA. In NRK-52E cells, PA-enhanced EV release was associated with an induction of cell apoptosis reflected by an increase in cleaved caspase-3 protein by Western blot and Annexin V positive cells analyzed by flow cytometry. Additionally, confocal microscopy confirmed the uptake of lipid-induced EVs by recipient renal proximal tubular cells. Collectively, our results indicate that PA stimulates EV release from cultured proximal tubular epithelial cells. Thus, extended characterization of lipid-induced EVs may constitute new signaling paradigms contributing to chronic kidney disease pathology.

Keyword: diabetes

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced insulin sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe insulin antagonism and body fat mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived fatty acids may impair insulin sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum fatty liver disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated fat-induced insulin resistance that defines in part the mechanisms of type 2 and nonalcoholic fatty liver disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of insulin-stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit insulin action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and fatty metabolism, insulin function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

Inhibition of insulin resistance by PGE1 via autophagy-dependent FGF21 pathway in diabetic nephropathy.

Insulin resistance is a critical process in the initiation and progression of diabetic nephropathy (DN). Alprostadil (Prostaglandin E1, PGE1) had protective effects on renal function. However, it is unknown whether PGE1 inhibited insulin resistance in renal tubule epithelial cells via autophagy, which plays a protective role in DN against insulin resistance. Insulin resistance was induced by (PA) in human HK-2 cells, shown as the decrease of insulin-stimulated AKT phosphorylation, glucose transporter-4 (GLUT4), glucose uptake and enhanced phosphorylation of insulin receptor substrate 1(IRS-1) at site serine 307 (pIRS-1ser307) and downregulated expression of IRS-1. Along with less abundance of p62, autophagy markers LC3B and Beclin-1 significantly increased in HK-2 cells exposed to PA. Such abnormal changes were significantly reversed by PGE1, which mimicked the role of autophagy gene 7 small interfering RNA (ATG7 siRNA). Furthermore, PGE1 promoted the protein expression of autophagy-related fibroblast growth factor-21 (FGF21), which alleviated insulin resistance. Results from western blotting and immunohistochemistry indicated that PGE1 remarkably restored autophagy, insulin resistance and the FGF21 expression in rat kidney of type 2 (T2DM). Collectively, we demonstrated the potential protection of PGE1 on insulin resistance in renal tubules via autophagy-dependent FGF21 pathway in preventing the progression of DN.

Keyword: diabetes

Palmitate and Stearate are Increased in the Plasma in a 6-OHDA Model of Parkinson\'s Disease.

Parkinson\'s disease (PD) is the second most common neurodegenerative disorder, without any widely available curative therapy. Metabolomics is a powerful tool which can be used to identify unexpected pathway-related disease progression and pathophysiological mechanisms. In this study, metabolomics in brain, plasma and liver was investigated in an experimental PD model, to discover small molecules that are associated with dopaminergic cell loss.Sprague Dawley (SD) rats were injected unilaterally with 6-hydroxydopamine (6-OHDA) or saline for the vehicle control group into the medial forebrain bundle (MFB) to induce loss of dopaminergic neurons in the substantia nigra pars compacta. Plasma, midbrain and liver samples were collected for metabolic profiling. Multivariate and univariate analyses revealed metabolites that were altered in the PD group.In plasma, ( = 3.72 × 10, FC = 1.81) and stearic ( = 3.84 × 10, FC = 2.15), were found to be increased in the PD group. ( = 3.5 × 10) and stearic ( = 2.7 × 10) correlated with test scores indicative of motor dysfunction. Monopalmitin ( = 4.8 × 10, FC = -11.7), monostearin ( = 3.72 × 10, FC = -15.1) and myo-inositol ( = 3.81 × 10, FC = -3.32), were reduced in the midbrain. The liver did not have altered levels of these molecules.Our results show that saturated free fatty acids, their monoglycerides and myo-inositol metabolism in the midbrain and enteric circulation are associated with 6-OHDA-induced PD pathology.

Keyword: diabetes

Sodium Orthovanadate Changes Fatty Composition and Increased Expression of Stearoyl-Coenzyme A Desaturase in THP-1 Macrophages.

Vanadium compounds are promising antidiabetic agents. In addition to regulating glucose metabolism, they also alter lipid metabolism. Due to the clear association between and atherosclerosis, the purpose of the present study was to assess the effect of sodium orthovanadate on the amount of individual fatty acids and the expression of stearoyl-coenzyme A desaturase (SCD or Δ-desaturase), Δ-desaturase, and Δ-desaturase in macrophages. THP-1 macrophages differentiated with phorbol 12-myristate 13-acetate (PMA) were incubated in vitro for 48\xa0h with 1\xa0μM or 10\xa0μM sodium orthovanadate (NaVO). The estimation of fatty composition was performed by gas chromatography. Expressions of the genes SCD, fatty desaturase 1 (FADS1), and fatty desaturase 2 (FADS2) were tested by qRT-PCR. Sodium orthovanadate in THP-1 macrophages increased the amount of saturated fatty acids (SFA) such as and stearic , as well as monounsaturated fatty acids (MUFA)-oleic and palmitoleic . Sodium orthovanadate caused an upregulation of SCD expression. Sodium orthovanadate at the given concentrations did not affect the amount of polyunsaturated fatty acids (PUFA) such as linoleic , arachidonic , eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic (DHA). In conclusion, sodium orthovanadate changed SFA and MUFA composition in THP-1 macrophages and increased expression of SCD. Sodium orthovanadate did not affect the amount of any PUFA. This was associated with a lack of influence on the expression of FADS1 and FADS2.

Keyword: diabetes

Docosahexaenoic antagonizes the boosting effect of on LPS inflammatory signaling by inhibiting gene transcription and ceramide synthesis.

It is well known that saturated fatty acids (SFAs) and unsaturated fatty , in particular omega-3 polyunsaturated fatty acids (n-3 PUFAs), have different effects on inflammatory signaling: SFAs are pro-inflammatory but n-3 PUFAs have strong anti-inflammatory properties. We have reported that (PA), a saturated fatty , robustly amplifies lipopolysaccharide (LPS) signaling to upregulate proinflammatory gene expression in macrophages. We also reported that the increased production of ceramide (CER) via sphingomyelin (SM) hydrolysis and CER de novo synthesis plays a key role in the synergistic effect of LPS and PA on proinflammatory gene expression. However, it remains unclear if n-3 PUFAs are capable of antagonizing the synergistic effect of LPS and PA on gene expression and CER production. In this study, we employed the above macrophage culture system and lipidomical analysis to assess the effect of n-3 PUFAs on proinflammatory gene expression and CER production stimulated by LPS and PA. Results showed that DHA strongly inhibited the synergistic effect of LPS and PA on proinflammatory gene expression by targeting nuclear factor kappa B (NFκB)-dependent gene transcription. Results also showed that DHA inhibited the cooperative effect of LPS and PA on CER production by targeting CER de novo synthesis, but not SM hydrolysis. Furthermore, results showed that myriocin, a specific inhibitor of serine palmitoyltransferase, strongly inhibited both LPS-PA-stimulated CER synthesis and proinflammatory gene expression, indicating that CER synthesis is associated with proinflammatory gene expression and that inhibition of CER synthesis contributes to DHA-inhibited proinflammatory gene expression. Taken together, this study demonstrates that DHA antagonizes the boosting effect of PA on LPS signaling on proinflammatory gene expression by targeting both NFκB-dependent transcription and CER de novo synthesis in macrophages.

Keyword: diabetes

Salvianolic B Attenuates Apoptosis of HUVEC Cells Treated with High Glucose or High Fat via Sirt1 Activation.

High glucose and high fat are important inducements for the development and progression of diabetic cardiopathy. Salvianolic B (SAB), which is the most abundant and bioactive compound in Danshen, attenuates oxidative stress-related disorders, such as cardiovascular diseases, cerebral ischemia, and . However, the effect of SAB on diabetic cardiopathy is not clear. The aim of study was to investigate the effect and the underlying molecular mechanisms of SAB on diabetic cardiopathy in vitro model. The human umbilical vein endothelial (HUVEC) cells were treated with high glucose (HG, 30 mM) or high fat (, PA, 0.75 mM) in the presence or absence of SAB (100, 200, and 400 mg/L) and incubated for 24 h. We found that HG or PA induced apoptosis of HUVEC cells, while treatment with SAB inhibited the apoptosis. We also found that SAB reversed HG- or PA-induced oxidative stress, apoptosis cell cytokines production, and expression of thioredoxin-interacting protein (TXNIP). Moreover, SAB increased HG- or PA-induced expression of Sirtuin 1 (Sirt1), a nicotinamide adenine dinucleotide- (NAD-) dependent histone deacetylase. Exposure of HUVEC cells to Ex527 (Sirt1 inhibitor) suppressed the effect of SAB on acetyl-p53 and procaspase-3 expressions. In conclusion, the results suggested that SAB could attenuate HUVEC cells damage treated with HG or PA via Sirt1 and might be a potential therapy agent for the diabetic cardiopathy treatment.

Keyword: diabetes

Plasma fatty acids in de novo lipogenesis pathway are associated with diabetogenic indicators among adults: NHANES 2003-2004.

Insulin regulates fatty acids (FAs) in the blood; conversely, FAs may mediate insulin sensitivity and are potentially modifiable risk factors of the diabetogenic state.The objective of our study was to examine the associations between plasma concentrations of FAs, fasting plasma glucose (FPG), and glycated hemoglobin (HbA1c) among individuals (n\xa0=\xa01433) in the NHANES (2003-2004).Plasma concentrations of 24 individual FAs were considered individually and in subgroups, per chemical structure. Study participants were categorized in diabetogenic groups: Group 1 (HbA1c ≥6.5% or FPG ≥126 mg/dL), Group 2 (HbA1c 5.7% to <6.5% or FPG 100 to <126 mg/dL), and Group 3 (HbA1c <5.7% and FPG <100 mg/dL). We assessed associations between diabetogenic groups and plasma FAs in multivariate multinomial regressions (with Group 3 as the reference).Overall, 7.0% of study participants were in Group 1; 33.3% were in Group 2. Plasma concentrations of several individual FAs, including even-chain saturated FAs (SFAs; myristic, , stearic acids) and monounsaturated FAs (MUFAs; cis-vaccenic, oleic acids), were respectively associated with greater odds of Groups 1 and 2 status, adjusting for covariates. Higher concentrations of SFA and MUFA subgroups (highest compared with lowest quartile) were associated with increased odds of Group 2 status [SFAs adjusted OR (aOR): 1.51 (95% CI: 1.05, 2.18); MUFAs aOR: 1.78 (95% CI: 1.11, 2.85)]. Higher eicosapentaenoic plasma concentration was associated with decreased odds of Group 1 status [quartile 4 aOR: 0.41 (95% CI: 0.17, 0.95)].Higher plasma concentrations of SFAs and MUFAs, primary de novo lipogenesis products, were associated with elevated FPG and HbA1c in a nationally representative study population in the United States. Additional studies are necessary to elucidate potential causal relationships between FAs (from endogenous production and dietary consumption) and diabetogenic indicators, as well as clinical implications for managing and prediabetes.

Keyword: diabetes

Pterostilbene reverses mediated insulin resistance in HepG2 cells by reducing oxidative stress and triglyceride accumulation.

Insulin resistance (IR) is known to precede onset of type 2 and increased oxidative stress appears to be a deleterious factor leading to IR. In this study, we evaluated ability of pterostilbene (PTS), a methoxylated analogue of resveratrol and a known antioxidant, to reverse (PA)-mediated IR in HepG2 cells. PTS prevented reactive oxygen species (ROS) formation and subsequent oxidative lipid damage by reducing the expression of NADPH oxidase 3 (NOX3) in PA treated HepG2 cells. Hepatic glucose production was used as a measure of IR and PTS reversed PA-mediated increase in hepatic glucose production by reducing expression of genes coding for gluconeogenic enzymes namely glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate carboxylase (PC); and their transcription factors cAMP response element binding protein (CREB) and fork head class Box O (FOXO1) along with its coactivator peroxisome proliferator-activated receptor gamma co-activator-1 α (PGC1α). PTS reversed PA-mediated activation of c-Jun N-terminal kinase (JNK), which in turn altered insulin signalling pathway by phosphorylating IRS-1 at Ser 307, leading to inhibition of phosphorylation of Akt and GSK-3β. PTS also reduced PA-mediated lipid accumulation by reducing expression of transcription factors SREBP1c and PPARα. SREBP1c activates genes involved in fatty and triglyceride synthesis while PPARα activates CPT1, a rate limiting enzyme for controlling entry and oxidation of fatty acids into mitochondria. PTS, however, did not influence PA uptake confirmed by using BODIPY-labelled fluorescent C16 fatty analogue. Thus, our data provides a possible mechanistic explanation for reversal of PA-mediated IR in HepG2 cells.

Keyword: diabetes

and Oleic : The Yin and Yang of Fatty Acids in Type 2 .

Increased plasma non-esterified fatty acids (NEFAs) link obesity with insulin resistance and type 2 (T2DM). However, in contrast to the saturated FA (SFA) , the monounsaturated FA (MUFA) oleic elicits beneficial effects on insulin sensitivity, and the dietary :oleic ratio impacts risk in humans. Here we review recent mechanistic insights into the beneficial effects of oleic compared with on insulin resistance and T2DM, including its anti-inflammatory actions, and its capacity to inhibit endoplasmic reticulum (ER) stress, prevent attenuation of the insulin signaling pathway, and improve β cell survival. Understanding the molecular mechanisms of the antidiabetic effects of oleic may contribute to understanding the benefits of this FA in the prevention or delay of T2DM.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Circulating Linoleic is Associated with Improved Glucose Tolerance in Women after Gestational .

Women with previously diagnosed gestational (GDM) are at increased risk of type-2- (T2D). We aimed to establish links between glucose tolerance (GT) and serum fatty (FA) profile in the transition from GDM to T2D. Six years after GDM, 221 women were grouped as having normal GT (NGT), impaired GT (IGT), or T2D based on oral GT test results. Fasting serum FAs were profiled, anthropometric measures taken, and dietary intake determined. Linoleic (LA) was significantly higher in NGT women ( < 0.001) compared with IGT and T2D, and emerged as a strong predictor of low glucose and insulin levels, independently of BMI. Self-reported vegetable oil consumption correlated with LA serum levels and glucose levels. Delta-6-, delta-9-, and stearoyl-CoA-desaturase activities were associated with decreased GT, and delta-5-desaturase activities with increased GT. In a subgroup of women at high risk of , low LA and high levels were seen in those that developed T2D, with no differences in other FAs or metabolic measurements. Results suggest that proportions of LA and are of particular interest in the transition from GDM to T2D. Interconversions between individual FAs regulated by desaturases appear to be relevant to glucose metabolism.

Keyword: diabetes

Argirein alleviates vascular endothelial insulin resistance through suppressing the activation of Nox4-dependent O production in diabetic rats.

Insulin resistance in endothelial cells contributes to the development of cardiovascular disease in type 2 (T2DM). Therefore, there are great potential clinical implications in developing pharmacological interventions targeting endothelial insulin resistance. Our previous studies indicated that argirein which was developed by combining rhein with L-arginine by a hydrogen bond, could substantially relieved stress related exacerbation of cardiac failure and alleviated cardiac dysfunction in T2DM, which was associated with suppressing NADPH oxidase activity. However, it is unclear whether argirein treatment attenuates the vascular lesion and dysfunction in T2DM and its underlying mechanisms.The rat aortic endothelial cells (RAECs) were used to treat with (PA), a most common saturated free fatty , which could induce insulin resistance. It was showed that argirein increased glucose uptake and glucose transporter-4 (Glut4) expression and reversed the phosphorylation of IRS-1-ser307 and AKT-ser473, consequently resulting in the increase of the production of eNOS and NO in PA-induced RAECs. We further found that argirein blocked the Nox4-dependent superoxide (O) generation, which regulated glucose metabolism in RAECs during PA stimulation. In vitro, argirein increased the release of endothelial NO to relieve the vasodilatory response to acetylcholine and insulin, and restored the expression of Nox4 and pIRS-1-ser307 in the aorta endothelium of high-fat diet (HFD)-fed rats following an injection of streptozocin (STZ).These results suggested that argirein could improve endothelial insulin resistance which was attributed to inhibiting Nox4-dependent redox signaling in RAECs. These studies thus revealed the novel effect of argirein to prevent the vascular complication in T2DM.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Metabolic modulation predicts heart failure tests performance.

The metabolic changes that accompany changes in Cardiopulmonary testing (CPET) and heart failure biomarkers (HFbio) are not well known. We undertook metabolomic and lipidomic phenotyping of a cohort of heart failure (HF) patients and utilized Multiple Regression Analysis (MRA) to identify associations to CPET and HFBio test performance (peak oxygen consumption (Peak VO2), oxygen uptake efficiency slope (OUES), exercise duration, and minute ventilation-carbon dioxide production slope (VE/VCO2 slope), as well as the established HF biomarkers of inflammation C-reactive protein (CRP), beta-galactoside-binding protein (galectin-3), and N-terminal prohormone of brain natriuretic peptide (NT-proBNP)). A cohort of 49 patients with a left ventricular ejection fraction < 50%, predominantly males African American, presenting a high frequency of , hyperlipidemia, and hypertension were used in the study. MRA revealed that metabolic models for VE/VCO2 and Peak VO2 were the most fitted models, and the highest predictors\' coefficients were from Acylcarnitine C18:2, , citric , asparagine, and 3-hydroxybutiric . Metabolic Pathway Analysis (MetPA) used predictors to identify the most relevant metabolic pathways associated to the study, aminoacyl-tRNA and amino biosynthesis, amino metabolism, nitrogen metabolism, pantothenate and CoA biosynthesis, sphingolipid and glycerolipid metabolism, fatty biosynthesis, glutathione metabolism, and pentose phosphate pathway (PPP). Metabolite Set Enrichment Analysis (MSEA) found associations of our findings with pre-existing biological knowledge from studies of human plasma metabolism as brain dysfunction and enzyme deficiencies associated with lactic acidosis. Our results indicate a profile of oxidative stress, lactic acidosis, and metabolic syndrome coupled with mitochondria dysfunction in patients with HF tests poor performance. The insights resulting from this study coincides with what has previously been discussed in existing literature thereby supporting the validity of our findings while at the same time characterizing the metabolic underpinning of CPET and HFBio.

Keyword: diabetes

Increased triacylglycerol - Fatty substrate cycling in human skeletal muscle cells exposed to eicosapentaenoic .

It has previously been shown that pretreatment of differentiated human skeletal muscle cells (myotubes) with eicosapentaenoic (EPA) promoted increased uptake of fatty acids and increased triacylglycerol accumulation, compared to pretreatment with oleic (OA) and (PA). The aim of the present study was to examine whether EPA could affect substrate cycling in human skeletal muscle cells by altering lipolysis rate of intracellular TAG and re-esterification of fatty acids. Fatty metabolism was studied in human myotubes using a mixture of fatty acids, consisting of radiolabelled oleic as tracer (14C-OA) together with EPA or PA. Co-incubation of myotubes with EPA increased cell-accumulation and incomplete fatty oxidation of 14C-OA compared to co-incubation with PA. Lipid distribution showed higher incorporation of 14C-OA into all cellular lipids after co-incubation with EPA relative to PA, with most markedly increases (3 to 4-fold) for diacylglycerol and triacylglycerol. Further, the increases in cellular lipids after co-incubation with EPA were accompanied by higher lipolysis and fatty re-esterification rate. Correspondingly, basal respiration, proton leak and maximal respiration were significantly increased in cells exposed to EPA compared to PA. Microarray and Gene Ontology (GO) enrichment analysis showed that EPA, related to PA, significantly changed i.e. the GO terms "Neutral lipid metabolic process" and "Regulation of lipid storage". Finally, an inhibitor of diacylglycerol acyltransferase 1 decreased the effect of EPA to promote fatty accumulation. In conclusion, incubation of human myotubes with EPA, compared to PA, increased processes of fatty turnover and oxidation suggesting that EPA may activate futile substrate cycling of fatty acids in human myotubes. Increased TAG-FA cycling may be involved in the potentially favourable effects of long-chain polyunsaturated n-3 fatty acids on skeletal muscle and whole-body energy metabolism.

Keyword: diabetes

Blood Fatty Profiles: New Biomarkers for Cardiometabolic Disease Risk.

Fatty (FA) profiles in different blood compartments are reflections of both diet and metabolism, and some FA levels are related to disease risk.Perhaps the most studied FA-disease relationship is between long-chain omega-3 polyunsaturated fatty acids [eicosapentaenoic (EPA) and docosahexaenoic (DHA)] and cardiovascular disease (CVD). Despite null results from recent large omega-3 FA supplementation trials, new research continues to support past studies showing that blood levels of EPA\u2009+\u2009DHA are inversely related to risk for total mortality and fatal CVD events. But blood levels of other FAs may also be useful markers of risk for a variety of diseases. The essential omega-6 FA linoleic is inversely associated with risk for developing type 2 (T2D), whereas risk for T2D is directly related to biomarkers of de novo lipogenesis ( and palmitoleic acids). Levels of industrially produced trans FAs have been linked to higher risk for CVD. Thus, blood levels of several individual FAs are emerging as modifiable biomarkers for risk of major chronic diseases.

Keyword: diabetes

-Induced Long Noncoding RNA Dnm3os Regulates Macrophage Functions and Inflammation via Nuclear Mechanisms.

Objective- Macrophages play key roles in inflammation and diabetic vascular complications. Emerging evidence implicates long noncoding RNAs in inflammation, but their role in macrophage dysfunction associated with inflammatory diabetic complications is unclear and was therefore investigated in this study. Approach and Results- RNA-sequencing and real-time quantitative PCR demonstrated that a long noncoding RNA Dnm3os (dynamin 3 opposite strand) is upregulated in bone marrow-derived macrophages from type 2 diabetic db/db mice, diet-induced insulin-resistant mice, and diabetic ApoE mice, as well as in monocytes from type 2 diabetic patients relative to controls. Diabetic conditions (high glucose and ) induced Dnm3os in mouse and human macrophages. Promoter reporter analysis and chromatin immunoprecipitation assays demonstrated that diabetic conditions induce Dnm3os via NF-κB activation. RNA fluorescence in situ hybridization and real-time quantitative PCRs of subcellular fractions demonstrated nuclear localization and chromatin enrichment of Dnm3os in macrophages. Stable overexpression of Dnm3os in macrophages altered global histone modifications and upregulated inflammation and immune response genes and phagocytosis. Conversely, RNAi-mediated knockdown of Dnm3os attenuated these responses. RNA pull-down assays with macrophage nuclear lysates identified nucleolin and ILF-2 (interleukin enhancer-binding factor 2) as protein binding partners of Dnm3os, which was further confirmed by RNA fluorescence in situ hybridization immunofluorescence. Furthermore, nucleolin levels were decreased in diabetic conditions, and its knockdown enhanced Dnm3os-induced inflammatory gene expression and histone H3K9-acetylation at their promoters. Conclusions- These results demonstrate novel mechanisms involving upregulation of long noncoding RNA Dnm3os, disruption of its interaction with nucleolin, and epigenetic modifications at target genes that promote macrophage inflammatory phenotype in . The data could lead to long noncoding RNA-based therapies for inflammatory complications.

Keyword: diabetes

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in . Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: diabetes

Downregulation of miR-320 Alleviates Endoplasmic Reticulum Stress and Inflammatory Response in 3T3-L1 Adipocytes.

MicroRNAs serve important roles in the regulation of endoplasmic reticulum stress (ERs). This study aimed to investigate the role of microRNA-320 (miR-320) in the development of ERs and the inflammatory response in 3T3-L1 adipocytes.The adipose tissue expression levels of miR-320 and ERs markers (GRP78, GRP94, Derlin-1 and CHOP) and the serum concentration of inflammatory cytokines (TNF-α, NF-κB and IL-6) in obese patients were evaluated using quantitative real-time RT-PCR. The correlation of miR-320 with genes involved in ERs and inflammation was analyzed. The effects of miR-320 on ERs and inflammation were explored using mature 3T3-L1 adipocytes, which were pretreated with (PA).ERs markers and inflammatory cytokines were all upregulated in obese patients. Adipose tissue miR-320 expression was also increased in obese patients, and had positive correlations with the levels of ERs markers and inflammatory cytokines. After PA treatment, the levels of ERs markers and inflammatory cytokines were elevated significantly in 3T3-L1 adipocytes. Moreover, miR-320 expression was increased in the cells under ERs status. The upregulation of miR-320 could enhance the expression of ERs markers and inflammatory cytokines, but the downregulation of miR-320 resulted in the opposite results.The data of this study indicate that miR-320 expression is upregulated in ERs status, and the downregulation of miR-320 ameliorates ERs and the inflammatory response in 3T3-L1 adipocytes. We consider that the approaches to decrease miR-320 expression may be novel therapeutic strategies for the treatment of obesity and obesity-related diseases.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

Type 2 Promotes Cell Centrosome Amplification via AKT-ROS-Dependent Signalling of ROCK1 and 14-3-3σ.

Type 2 is associated with oxidative stress and DNA damage which can cause centrosome amplification. Thus, the study investigated centrosome amplification in type 2 and the underlying mechanisms.Centrosome numbers in human peripheral blood mononuclear blood cells (PBMC) from healthy subjects and patients with type 2 were compared to access the association between type 2 and centrosome amplification. Colon cancer cells were used to investigate the molecular mechanisms underlying the centrosome amplification triggered by high glucose, insulin and . Western blot analysis was used to quantify the level of protein and protein phosphorylation. Immunofluorescent staining was performed to detect centrosomes. ROS was quantified using flow cytometry technique. Transcriptpmic profiling was performed using Illumina HiSeqTM500 platform.We found that centrosome amplification was increased PBMC from the type 2 diabetic patients, which correlated with the levels of fasting blood glucose and HbA1c. High glucose, insulin and , alone or in combinations, induced ROS production and centrosome amplification. Together, they increased AKT activation as well as the expression, binding and centrosome translation of ROCK1 and 14-3-3σ. Results from further analyses showed that AKT-ROS-dependent upregulations of expression, binding and centrosome translocation of ROCK1 and 14-3-3σ was the molecular pathway underlying the centrosome amplification in vitro triggered by high glucose, insulin and . Moreover, the key in vitro molecular signalling events activated by high glucose, insulin and were verified in PBMC from the patients with type 2 .Our results show that type 2 promotes cell centrosome amplification, and suggest that the diabetic pathophysiological factors-activated AKT-ROS-dependent signalling of ROCK1 and 14-3-3σ is the underlying molecular mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Role of glutamine synthetase in angiogenesis beyond glutamine synthesis.

Glutamine synthetase, encoded by the gene GLUL, is an enzyme that converts glutamate and ammonia to glutamine. It is expressed by endothelial cells, but surprisingly shows negligible glutamine-synthesizing activity\xa0in these cells at physiological glutamine levels. Here we show in mice that genetic deletion of Glul in endothelial cells impairs vessel sprouting during vascular development, whereas pharmacological blockade of glutamine synthetase suppresses angiogenesis in ocular and inflammatory skin disease while only minimally affecting healthy adult quiescent endothelial cells. This relies on the inhibition of endothelial cell migration but not proliferation. Mechanistically we show that in human umbilical vein endothelial cells GLUL knockdown reduces membrane localization and activation of the GTPase RHOJ while activating other Rho GTPases and Rho kinase, thereby inducing actin stress fibres and impeding endothelial cell motility. Inhibition of Rho kinase rescues the defect in endothelial cell migration that is induced by GLUL knockdown. Notably, glutamine synthetase palmitoylates itself and interacts with RHOJ to sustain RHOJ palmitoylation, membrane localization and activation. These findings reveal that, in addition to the\xa0known formation of glutamine, the enzyme glutamine synthetase shows unknown\xa0activity in endothelial cell migration during pathological angiogenesis\xa0through RHOJ palmitoylation.

Keyword: diabetes

Rare ginsenosides ameliorate lipid overload-induced myocardial insulin resistance via modulating metabolic flexibility.

Rare ginsenosides are found in ginseng and notoginseng, two medicinal plants widely used in China for treatment of cardiovascular diseases and type 2 . However, their pharmacological studies regarding myocardial fuel metabolism and insulin signaling are not clear.To explore the effect of a rare ginsenoside-standardized extract (RGSE), derived from steamed notoginseng, on cardiac fuel metabolism and insulin signaling.We used (PA) to treat H9c2 cells in vitro and high fat diet (HFD) to mice to induce insulin resistance in vivo.In vitro, differentiated H9c2 cells were pretreated with RGSE, metformin, mildronate or dichloroacetate (DCA) and stimulated with PA. In vivo, mice were fed with HFD and received RGSE, metformin or DCA for 6 weeks. Protein expression was determined by Western blotting. Mitochondrial membrane potential (Δψm), glucose uptake and reactive oxygen species (ROS) production were measured by fluorescence labeling. Other assessments including oxygen consumption rate (OCR) were also performed.RGSE prevented PA-induced decrease in pyruvate dehydrogenase (PDH) activity and increase in carnitine palmitoyltransferase 1 (CPT1) expression, and ameliorated insulin-mediated glucose uptake and utilization in H9c2 cells. Metformin and mildronate exhibited similar effects. In vivo, RGSE counteracted HFD-induced increase in myocardial expression of p-PDH and CPT1 and ameliorated cardiac insulin signaling. Metformin and DCA also showed beneficial effects. Further study showed that RGSE decreased OCR and mitochondrial complex I activity in PA-treated H9c2 cells, reduced ROS production and relieved mitochondrial oxidative stress, thus decreased serine phosphorylation in IRS-1.RGSE ameliorated myocardial insulin sensitivity under conditions of lipid overload, which was tightly associated with the decrease in mitochondrial oxidative stress via modulating glucose and fatty oxidation.Copyright © 2018. Published by Elsevier GmbH.

Keyword: diabetes

Metabolomics analysis of the protective effect of rubusoside on -induced lipotoxicity in INS-1 cells using UPLC-Q/TOF MS.

is one of the most severe chronic diseases worldwide. It is widely accepted that apoptosis of the pancreatic beta cell is an important cause for the induction of hyperglycemia and high levels of free fatty acids (FFAs), also called lipotoxicity associated with pancreatic beta cell dysfunction. Lipotoxicity has been proven to be an important pathogenic factor of . However, until now, the mechanism of FFA-induced lipotoxicity in INS-1 cells has not been fully understood. Current anti-diabetic drugs that protect islet cells are often toxic to healthy cells, resulting in negative side effects. Thus, there is an urgent need to identify more effective and safer anti-diabetic agents to protect pancreatic islet cells. Rubusoside (RUB) is a major ingredient in the leaves of Rubus suavissimus S. Lee, which decreases blood glucose levels by protecting pancreatic islet cells. However, the exact mechanism of this effect is unknown. In this study, metabolomics experiments based on UPLC-Q/TOF MS characterized a total of 15 metabolites as potential biomarkers associated with lipotoxicity induced by in INS-1 cells. According to the metabolic pathway analysis, pentose and glucuronate interconversions, and glycerophospholipid metabolism were recognized as the most influenced metabolic pathways associated with lipotoxicity. Unexpectedly, deviations of 14 metabolites in lipotoxic INS-1 cells were regulated by RUB, suggesting synergistic mediation of the abnormal metabolic pathways. The metabolomics strategy based on UPLC/Q-TOF MS analysis provides a new insight into the mechanisms of lipotoxicity induced by and the anti-lipotoxic activity of RUB in INS-1 cells.

Keyword: diabetes

The role and mechanism of K3.1 channels in human monocyte migration induced by .

Monocyte migration into diseased tissues contributes to the pathogenesis of diseases. Intermediate-conductance Ca-activated K (K3.1) channels play an important role in cell migration. However, the role of K3.1 channels in mediating monocyte migration induced by (PA) is still unclear. Using cultured THP-1 cells and peripheral blood mononuclear cells from healthy subjects, we investigated the role and signaling mechanisms of K3.1 channels in mediating the migration induced by PA. Using methods of Western blotting analysis, RNA interference, cell migration assay and ELISA, we found that PA-treated monocytes exhibited increment of the protein levels of K3.1 channel and monocyte chemoattractant protein-1 (MCP-1), and the effects were reversed by co-incubation of PA with anti-TLR2/4 antibodies or by specific inhibitors of p38-MAPK, or NF-κB. In addition, PA increased monocyte migration, which was abolished by a specific K3.1 channel blocker, TRAM-34, or K3.1 small interfering RNA (siRNA). The expression and secretion of MCP-1 induced by PA was also similarly prevented by TRAM-34 and K3.1 siRNA. These results demonstrate for the first time that PA upregulates K3.1 channels through TLR2/4, p38-MAPK and NF-κB pathway to promote the expression of MCP-1, and then induce the trans-endothelial migration of monocytes.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Permethrin and ivermectin modulate lipid metabolism in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 , which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid metabolism. The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates fatty oxidation. Results in this study suggested that hepatic lipid metabolism may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 and Obesity in the Rat, as Suggested by the Effects on Stem Cell Culture.

Previous work showed that muscle-derived stem cells (MDSCs) exposed long-term to the milieu of uncontrolled type 2 (UC-T2D) in male obese Zucker (OZ) rats, were unable to correct the associated erectile dysfunction and the underlying histopathology when implanted into the corpora cavernosa, and were also imprinted with a noxious gene global transcriptional signature (gene-GTS), suggesting that this may interfere with their use as autografts in stem cell therapy.To ascertain the respective contributions of dyslipidemia and hyperglycemia to this MDSC damage, clarify its mechanism, and design a bioassay to identify the damaged stem cells.Early MDSCs and late MDSCs were respectively isolated from nearly normal young OZ rats and moderately hyperglycemic and severely dyslipidemic/obese aged rats with erectile dysfunction. Monolayer cultures of early diabetic MDSCs were incubated 4 days in DMEM/10% fetal calf serum\xa0+ or\xa0- aged OZ or lean Zucker serum from non-diabetic lean Zucker rats (0.5-5%) or with soluble (PA) (0.5-2 mM), cholesterol (CHOL) (50-400 mg/dL), or glucose (10-25 mM).Fat infiltration was estimated by Oil red O, apoptosis by TUNEL, protein expression by Western blots, and gene-GTS and microRNA (miR)-GTS were determined in these stem cells\' RNA.Aged OZ serum caused fat infiltration, apoptosis, myostatin overexpression, and impaired differentiation. Some of these changes, and also a proliferation decrease occurred with PA and CHOL. The gene-GTS changes by OZ serum did not resemble the in\xa0vivo changes, but some occurred with PA and CHOL. The miR-GTS changes by OZ serum, PA, and CHOL resembled most of the in\xa0vivo changes. Hyperglycemia did not replicate most alterations.MDSCs may be damaged in long-term UC-T2D/obese patients and be ineffective in autologous human stem cell therapy, which may be prevented by excluding the damaged MDSCs.The in\xa0vitro test of MDSCs is innovative and fast to define dyslipidemic factors inducing stem cell damage, its mechanism, prevention, and counteraction. Confirmation is required in other T2D/obesity rat models and stem cells (including human), as well as miR-GTS biomarker validation as a stem cell damage biomarker.Serum from long-term UC-T2D/obese rats or dyslipidemic factors induces a noxious phenotype and miR-GTS on normal MDSCs, which may lead in\xa0vivo to the repair inefficacy of late diabetic MDSCs. This suggests that autograft therapy with MDSCs in long-term UT-T2D obese patients may be ineffective, albeit this may be predictable by prior stem cell miR-GTS tests. Masouminia M, Gelfand R, Kovanecz I, et\xa0al. Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 and Obesity in the Rat, as Suggested by the Effects on Stem Cell Culture. J Sex Med 2018;15:1678-1697.Copyright © 2018 International Society for Sexual Medicine. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

A prospective and longitudinal study of plasma phospholipid saturated fatty profile in relation to cardiometabolic biomarkers and the risk of gestational .

Data on saturated fatty acids (SFAs) in relation to metabolic function and glucose homeostasis remain controversial. Such data are lacking among pregnant women.We prospectively investigated objectively measured individual and subclasses of plasma phospholipid SFAs throughout pregnancy in relation to cardiometabolic markers and gestational (GDM) risk.Within the National Institute of Child Health and Human Development Fetal Growth Studies-Singleton Cohort of 2802 singleton pregnancies, 107 GDM cases were ascertained via medical record review and matched to 214 non-GDM controls on age, race/ethnicity, and gestational week (GW) at blood collection. Individual plasma phospholipid SFA concentrations were repeatedly measured throughout pregnancy at GWs 10-14, 15-26, 23-31, and 33-39 and also grouped into subclasses of even- or odd-chain SFAs.From GW 10, even-chain SFA concentrations were significantly higher among women who later developed GDM, whereas odd-chain SFAs were significantly lower among GDM cases compared with controls. At GWs 10-14, the SFA (16:0) was positively associated with impaired insulin resistance and cardiometabolic markers and the risk of GDM [adjusted OR comparing the highest with the lowest quartile (aORQ4-Q1): 4.76; 95% CI: 1.72, 13.10; P-trend\xa0=\xa00.001]. In contrast, odd-chain SFAs were inversely related to the previously mentioned markers and GDM risk [aORQ4-Q1 for pentadecanoic (15:0): 0.32; 95% CI: 0.11, 0.92; P-trend\xa0=\xa00.025; for heptadecanoic (17:0): 0.20; 95% CI: 0.07, 0.58; P-trend\xa0=\xa00.003]. Women with high (median or greater) even-chain SFA concentrations and low (less than median) odd-chain SFAs had a 9.43-fold (95%: CI 3.26-, 27.30-fold) increased risk compared with women with low even-chain and high odd-chain SFA concentrations. Similar results were observed at GWs 15-26.The study provided one of the first lines of evidence suggesting that circulating concentrations of SFAs varying by SFA chain length, as early as GWs 10-14, were significantly and differentially associated with subsequent risk of GDM. Our findings highlight the importance of assessing objectively measured, individual, and subclasses of SFAs to investigate their distinct biological and pathophysiologic roles in glucose homeostasis and cardiometabolic outcomes. This study was registered at www.clinicaltrials.gov as .

Keyword: diabetes

Sirtuin 5 overexpression attenuates glucolipotoxicity-induced pancreatic β cells apoptosis and dysfunction.

Recently, SIRT5 was reported to be a predominant desuccinylase and demalonylase in mitochondria. Ablation of SIRT5 enhances the systemic succinylation and malonylation of mitochondrial proteins, including various metabolic enzymes; however, its function in pancreatic β cells has not yet been clarified. In this study, we evaluated the effects of SIRT5 overexpression on glucolipotoxicity-induced apoptosis in β cell lines. Full-length SIRT5, which preferentially targeted to mitochondria and partially to the nucleus and cytoplasm, was overexpressed in NIT-1 cells. Chronic exposure to excess palmitate and glucose (High-PA-G) induced apoptosis and suppressed glucose-stimulated insulin secretion in β cells. SIRT5 overexpression significantly alleviated apoptosis under the High-PA-G condition, accompanied by suppressed Caspase 3 activity and reduced malondialdehyde levels. SIRT5 overexpression also improved β cell secretory capacity in response to glucose under the High-PA-G condition, suggesting its protective role in β cell function. Furthermore, SIRT5 overexpression reversed the decreasing trend of anti-apoptotic factors BCL-2 and BCL-XL expression under High-PA-G condition. Further regulation mechanisms between SIRT5 and these anti-apoptotic factors remains to be explored in future studies. Our data reveal that SIRT5 is a potentially protective factor for pancreatic β cells against glucolipotoxicity-induced apoptosis and cell dysfunction.Copyright © 2018. Published by Elsevier Inc.

Keyword: diabetes

Postprandial Saturated Fatty Acids Increase the Risk of Type 2 : A Cohort Study in a Chinese Population.

Experimental evidence suggests saturated fatty acids (SFAs) are associated with insulin resistance, but results from epidemiological studies on fasting SFAs- risk are inconsistent.We investigated SFA (fasting and 2-hour postprandial) profiles and risk.A total of 8940 participants were recruited for the Harbin People\'s Health Study in 2008. Serum SFAs (fasting and 2-hour postprandial) at baseline in Chinese men and women without were profiled, and type 2 was ascertained using World Health Organization criteria after 4 to 7 years of follow-up.Associations between 2-hour postprandial SFA (2h-SFA) and .At baseline, incident cases of were older with a higher body mass index and waist circumference. After a mean follow-up of 6.7 years, 658 incident cases of occurred. After propensity score computation and inverse probability of treatment weighting (IPTW) estimation, fasting SFAs were unrelated to risk but IPTW-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for the highest tertile of 2-hour postprandial stearic (2h-SA), 2-hour postprandial (2h-PA), and 2h-SFA for risk were 2.50 (2.08 to 3.16), 1.56 (1.23 to 2.02), and 1.70 (1.34 to 2.17), respectively (P-trend < 0.0001). Similarly, 2h-SA/fasting SA, 2h-PA/fasting PA, and 2h-SFA/fasting SFA ratios [IPTW-adjusted OR (95% CI): 2.94 (2.39 to 3.58), 2.31 (1.80 to 2.93), and 2.42 (1.91 to 3.11), respectively; P-trend < 0.0001] predicted the risk.Higher serum 2h-SFA (but not fasting SFA) independently predicted risk.

Keyword: diabetes

Changes of MODY signal pathway genes in the endoplasmic reticulum stress in INS-1-3 cells.

Metabolic disturbances induce endoplasmic reticulum stress (ERS) in pancreatic beta cells. This study aims to investigate whether a common pathway exists in the ERS induced by various chemicals, including high levels of glucose and palmitate in INS-1-3 cells.ERS in INS-1-3 cells was induced by exposure cells to thapsigargin (TG), tunicamycin (TM) or (PA) +high glucose (HG). Digital gene expression (DGE) profiling technique was used to detect differentially expressed genes. The profile of gene expression was detected by gene oncology (GO) function and pathway enrichment analysis. Nkx6.1 over-expression was established in INS-1-3 cell lines by lentivirus infection to revert the inhibition of Nkx6.1 expression found in the situation of ERS. Real time reverse transcription polymerase chain reaction (RT-PCR) was used to verify the expression changes of key genes. Cell viability was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The apoptosis was determined by flow cytometry. INS-1-3 cell function was measured by glucose stimulated insulin secretion test(GSIS).As compared to control, DGE demonstrated that there were 135, 57 and 74 differentially expressed genes in TM, TG and HG+PA groups, respectively. Those differentially expressed genes were enriched to ERS, antigen processing and presentation, protein export pathways, and interestingly, the maturity onset of the young (MODY) pathway. Nkx6.1 is one of common down-regulated gene in MODY signaling pathway among TM, TG and HG+PA groups. Over-expression of Nkx6.1 ameliorated glucolipotoxicity induced apoptosis rate by 45.4%, and increased proliferation by 40.9%. At the same time, GSIS increased by 1.82 folds.MODY pathway genes expression was changed in the state of ERS. Over-expression of Nkx6.1 protected the INS-1-3 cells from glucolipotoxicity.

Keyword: diabetes

Rational design of dimeric lipidated Xenopus glucagon-like peptide 1 analogues as long-acting antihyperglycaemic agents.

Dimerization is viewed as an effective means to enhance the binding affinity and therapeutic potency of peptides. Both dimerization and lipidation effectively prolong the half-life of peptides in\xa0vivo by increasing hydrodynamic size and facilitating physical interactions with serum albumin. Here, we report a novel method to discover long-acting glucagon-like peptide 1 (GLP-1) analogues by rational design based on Xenopus GLP-1 through a combined dimerization and lipidization strategy. On the basis of our previous structure analysis of Xenopus GLP-1, and a C-terminal Cys were firstly introduced into two Xenopus GLP-1 analogues (1 and 2), and the afforded 3 and 4 were further reacted with bis-maleimide amine to afford two dimeric lipidated Xenopus GLP-1 analogues (5 and 6). The in\xa0vitro and in\xa0vivo biological activities of 5 and 6 were significantly improved as compared with their monomers. Moreover, the selected compound 6 showed greater hypoglycemic and insulinotropic activities than liraglutide even when the dose of 6 was reduced to half in db/db mice. Pharmacokinetic test revealed that 6 had a ∼ 3-fold longer half-life than liraglutide in Kunming mice and SD rats, and the longer half-life of 6 led to excellent long-acting hypoglycemic effects as confirmed by two different pharmacological methods conducted on db/db mice. Finally, a 7 weeks chronic study conducted on db/db mice demonstrated the better therapeutic efficacies of 6 on glucose tolerance normalization, HbA1c reduction and pancreas islets protection than liraglutide. The present research showed that combined dimerization and lipidization is effective when applied to Xenopus GLP-1 analogue to develop novel GLP-1 analogue for the treatment of type 2 . In addition, the promising preclinical data of 6 suggested the therapeutic potential of 6 as a novel anti-diabetic agent.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Excessive Autophagy Activation and Increased Apoptosis Are Associated with -Induced Cardiomyocyte Insulin Resistance.

Diabetic cardiomyopathy (DCM) remains the major cause of death associated with . Researchers have demonstrated the importance of impaired cardiac insulin signaling in this process. Insulin resistance (IR) is an important predictor of DCM. Previous studies examining the dynamic changes in autophagy during IR have yielded inconsistent results. This study aimed to investigate the dynamic changes in autophagy and apoptosis in the rat H9c2 cardiomyocyte IR model. H9c2 cells were treated with 500\u2009M (PA) for 24 hours, resulting in the induction of IR. To examine autophagy, monodansylcadaverine staining, GFP-LC3 puncta confocal observation, and Western blot analysis of LC3I-to-LC3II conversion were used. Results of these studies showed that autophagic vesicles increased in numbers during the first 24 hours and then decreased by 36 hours after PA treatment. Western blot analysis showed that treatment of H9c2 cells with 500\u2009M PA for 24 hours decreased the expression of Atg12-Atg5, Atg16L1, Atg3, and PI3Kp85. Annexin V/PI flow cytometry revealed that PA exposure for 24 hours increased the rate of apoptosis. Together, this study demonstrates that PA induces IR in H9c2 cells and that this process is accompanied by excessive activation of autophagy and increases in apoptosis.

Keyword: diabetes

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Lipid Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial lipid metabolism.A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and lipid profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of -induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate -induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial lipid metabolism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

miR-125a-5p ameliorates hepatic glycolipid metabolism disorder in type 2 through targeting of STAT3.

Glycolipid metabolic disorder is an important cause for the development of type 2 (T2DM). Clarification of the molecular mechanism of metabolic disorder and exploration of drug targets are crucial for the treatment of T2DM. We examined miR-125a-5p levels in -induced AML12 cells and the livers of type 2 diabetic rats and mice, and then validated its target gene. Through gain- and loss-of-function studies, the effects of miR-125a-5p via targeting of STAT3 on regulating glycolipid metabolism were further illustrated and . We found that miR-125a-5p was significantly decreased in the livers of diabetic mice and rats, and STAT3 was identified as the target gene of miR-125a-5p. Overexpression of miR-125a-5p in C57BL/6 mice decreased STAT3 level and downregulated the expression levels of p-STAT3 and SOCS3. Consequently, SREBP-1c-mediated lipogenesis pathway was inhibited, and PI3K/AKT pathway was activated. Moreover, silencing of miR-125a-5p significantly increased the expression levels of STAT3, p-STAT3 and SOCS3, thus activating SREBP-1c pathway and suppressing PI3K/AKT pathway. Therefore, hyperglycemia, hyperlipidemia and decreased liver glycogen appeared in C57BL/6 mice. In -induced AML12 cells, miR-125a-5p mimic markedly increased glucose consumption and uptake and decreased the accumulation of lipid droplets by regulating STAT3 signaling pathway. Consistently, miR-125a-5p overexpression obviously inhibited STAT3 expression in diabetic KK-Ay mice, thereby decreasing blood glucose and lipid levels, increasing hepatic glycogen content, and decreasing accumulation of hepatic lipid droplets in diabetic mice. Furthermore, inhibition of miR-125a-5p in KK-Ay mice aggravated glycolipid metabolism dysfunction through regulating STAT3. Our results confirmed that miR-125a-5p should be considered as a regulator of glycolipid metabolism in T2DM, which can inhibit hepatic lipogenesis and gluconeogenesis and elevate glycogen synthesis by targeting STAT3.

Keyword: diabetes

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic fatty liver disease (NAFLD), and type 2 (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 lipid species were identified, quantified, and classified into free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of fatty acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

Keyword: diabetes

Impact of Brain Fatty Signaling on Peripheral Insulin Action in Mice.

Glucose homeostasis and energy balance are under control by peripheral and brain processes. Especially insulin signaling in the brain seems to impact whole body glucose homeostasis and interacts with fatty signaling. In humans circulating saturated fatty acids are negatively associated with brain insulin action while animal studies suggest both positive and negative interactions of fatty acids and insulin brain action. This apparent discrepancy might reflect a difference between acute and chronic fatty signaling. To address this question we investigated the acute effect of an intracerebroventricular administration on peripheral glucose homeostasis. We developed and implemented a method for simultaneous monitoring of brain activity and peripheral insulin action in freely moving mice by combining radiotelemetry electrocorticography (ECoG) and euglycemic-hyperinsulinemic clamps. This method allowed gaining insight in the early kinetics of brain fatty signaling and its contemporaneous effect on liver function , which, to our knowledge, has not been assessed so far in mice.Insulin-induced brain activity in the theta and beta band was decreased by acute intracerebroventricular application of . Peripherally it amplified insulin action as demonstrated by a significant inhibition of endogenous glucose production and increased glucose infusion rate. Moreover, our results further revealed that the brain effect of peripheral insulin is modulated by load in the brain.These findings suggest that insulin action is amplified in the periphery and attenuated in the brain by acute application. Thus, our results indicate that acute signaling in the brain may be different from chronic effects.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

Sonodynamic therapy inhibits palmitate-induced beta cell dysfunction via PINK1/Parkin-dependent mitophagy.

In type 2 (T2DM), the overload of glucose and lipids can promote oxidative stress and inflammatory responses and contribute to the failure of beta cells. However, therapies that can modulate the function of beta cells and thus prevent their failure have not been well explored. In this study, beta cell injury model was established with (PA) to simulate the lipotoxicity (high-fat diet) found in T2DM. Sonodynamic therapy (SDT), a novel physicochemical treatment, was applied to treat injured beta cells. We found that SDT had specific effects on mitochondria and induced transient large amount of mitochondrial reactive oxygen species (ROS) production in beta cells. SDT also improved the morphology and function of abnormal mitochondria, inhibited inflammatory response and reduced beta cell dysfunction. The improvement of mitochondria was mediated by PINK1/Parkin-dependent mitophagy. Additionally, SDT rescued the transcription of PINK1 mRNA which was blocked by PA treatment, thus providing abundant PINK1 for mitophagy. Moreover, SDT also increased insulin secretion from beta cells. The protective effects of SDT were abrogated when mitophagy was inhibited by cyclosporin A (CsA). In summary, SDT potently inhibits lipotoxicity-induced beta cell failure via PINK1/Parkin-dependent mitophagy, providing theoretical guidance for T2DM treatment in aspects of islet protection.

Keyword: diabetes

Theaflavins Improve Insulin Sensitivity through Regulating Mitochondrial Biosynthesis in -Induced HepG2 Cells.

Theaflavins, the characteristic and bioactive polyphenols in black tea, possess the potential improving effects on insulin resistance-associated metabolic abnormalities, including obesity and type 2 . However, the related molecular mechanisms are still unclear. In this research, we investigated the protective effects of theaflavins against insulin resistance in HepG2 cells induced by . Theaflavins significantly increased glucose uptake of insulin-resistant cells at noncytotoxic doses. This activity was mediated by upregulating the total and membrane bound glucose transporter 4 protein expressions, increasing the phosphor-Akt (Ser473) level, and decreasing the phosphorylation of IRS-1 at Ser307. Moreover, theaflavins were found to enhance the mitochondrial DNA copy number, down-regulate the PGC-1β mRNA level and increase the PRC mRNA expression. Mdivi-1, a selective mitochondrial division inhibitor, could attenuate TFs-induced promotion of glucose uptake in insulin-resistant HepG2 cells. Taken together, these results suggested that theaflavins could improve hepatocellular insulin resistance induced by free fatty acids, at least partly through promoting mitochondrial biogenesis. Theaflavins are promising functional food ingredients and medicines for improving insulin resistance-related disorders.

Keyword: diabetes

Nutritionally Derived Metabolic Cues Typical of the Obese Microenvironment Increase Cholesterol Efflux Capacity of Adipose Tissue Macrophages.

Cholesterol retention within plasma membranes of macrophages is associated with increased inflammatory signaling. Cholesterol efflux via the transporters ABCA1, ABCG1, and SR-BI to high-density lipoprotein (HDL) particles is a critical mechanism to maintain cellular cholesterol homeostasis. Little is known about the impact of the obese microenvironment on cholesterol efflux capacity (CEC) of macrophages. In this study, the CEC of obese-derived primary adipose-tissue macrophages (ATM) is evaluated and the in vivo microenvironment is modeled in vitro to determine mechanisms underlying modulated CEC.F4/80 ATM are labeled with H-cholesterol ex vivo, and CEC and ABCA1/ABCG1 protein levels are determined. Total, ABCA1-dependent, and ABCA1-independent CECs are determined in J774 macrophages polarized to M1 (LPS&IFNγ), M2 (IL-4&IL-13), or metabolic phenotypes (glucose, insulin, and ).Obese ATM exhibit enhanced CEC and ABCA1 and ABCG1 expression compared to lean ATM. In contrast, ABCA1-CEC is suppressed from M1 polarized macrophages compared to untreated in vitro, by activation of the JAK/STAT pathway. Incubation of macrophages in vitro in high glucose augments cAMP-induced ABCA1 protein expression and ABCA1-CEC.These novel findings demonstrate remarkable plasticity of macrophages to respond to their environment with specific modulation of ABCA1 depending on whether classical pro-inflammatory or metabolic cues predominate.© 2018 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: diabetes

Individual free fatty acids have unique associations with inflammatory biomarkers, insulin resistance and insulin secretion in healthy and gestational diabetic pregnant women.

We investigated the relationships of maternal circulating individual free fatty acids (FFA) with insulin resistance, insulin secretion and inflammatory biomarkers during mid-pregnancy.The data were drawn from a prospective cohort of generally healthy pregnant women (n=1368, African-American 36%, Hispanic 48%, Caucasian 16%) in Camden, NJ. We quantitatively determined 11 FFAs, seven cytokine/adipokine, homeostatic model assessment of insulin resistance (HOMA-IR) and C-peptide levels from the fasting blood samples that were collected at 16 weeks of gestation. Multivariate analyses were performed along with separate analyses for each individual FFA.High HOMA-IR (p<0.001) and C-peptide (p<0.0001) levels were positively associated with a twofold to fourfold increased risk for developing gestational (GDM). Negative relationships were found with specific FFAs (molecular percentage, palmitoleic, oleic, linolenic, myristic acids) and HOMA-IR and C-peptide levels (p<0.01\u2009to p<0.0001). In contrast, , stearic, arachidonic, dihomo-γ-linolenic (DGLA) and docosahexaenoic acids were positively associated with HOMA-IR and C-peptide (p<0.01\u2009to p<0.0001). The individual FFAs also predicted cytokine/adipokine levels. For example, women who had elevated DGLA (highest quartile) were twice as (adjusted OR 2.06, 95%\u2009CI 1.42 to 2.98) likely to have higher interleukin (IL)-8 (p<0.0001) levels. Conversely, women with high palmitoleic, oleic, and linolenic levels had reduced odds (≥2-fold, p<0.01\u2009to p<0.001) for having higher IL-8, IL-6 or tumor necrosis factor-alpha levels.Our results suggest that maternal individual FFAs uniquely affect insulin resistance and secretion. The effects are either direct or indirect via modulation of the inflammatory response. Modifying the composition of FFAs may help in reducing the risk of GDM.

Keyword: diabetes

Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by lipid metabolism-related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 . However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic fat accumulation in rats with high-fat diet (HFD) induced obesity. Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and insulin tolerance tests, and immunohistochemistry. The expression of lipid metabolism- and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on insulin release in co-cultures with isolated rat islets.HFD increased body weight and ectopic fat accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and insulin resistance, upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of lipid metabolism-related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and insulin secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of lipid metabolism, suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in . In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.

Keyword: diabetes

MicroRNA‑29a is involved lipid metabolism dysfunction and insulin resistance in C2C12 myotubes by targeting PPARδ.

MicroRNA‑29a (miR‑29a) expression has been reported to be closely associated with skeletal muscle insulin resistance and type 2 . The present study investigated the effect of miR‑29a on (PA)‑induced lipid metabolism dysfunction and insulin resistance in C2C12 myotubes via overexpressing or silencing of miR‑29a expression. Mouse C2C12 myoblasts were cultured, differentiated and transfected with miR‑29a or miR‑29a inhibitor lentiviral with or without subsequent (PA) treatment. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis were performed to assess the mRNA and protein levels of related genes, respectively. PA treatment increased the expression of miR‑29a in a time‑ and dose‑ dependent manner. miR‑29a silencing improved insulin‑induced glucose uptake and increased glucose transporter‑4 (GLUT4) transportation to the plasma membrane by upregulating its target peroxisome proliferator‑activated receptor δ (PPARδ). Furthermore, it was observed that miR‑29a regulated the expression of genes associated with lipid metabolism, including pyruvate dehydrogenase kinase isoform, mitochondrial uncoupling protein (UCP)2, UCP3, long chain specific acyl‑CoA dehydrogenase, mitochondrial and fatty transport protein 2. The results confirmed that silencing miR‑29a induced a decrease in glucose transport and affected lipid metabolism in PA‑treated C2C12 cells, and therefore may be involved in insulin resistance by targeting PPARδ in skeletal muscle. Therefore, the inhibition of miR‑29a may be a potential novel strategy for treating insulin resistance and type 2 .

Keyword: diabetes

HAMSCs/HBMSCs coculture system ameliorates osteogenesis and angiogenesis against glucolipotoxicity.

Osteoporosis and vascular lesions induced by glucolipotoxicity are common complications of (DM). In order to deal with these complications, we designed a new therapeutic strategy, i.e. coculture system containing human amnion-derived mesenchymal stem cells (HAMSCs) and human bone marrow mesenchymal stem cells (HBMSCs). Two in\xa0vitro coculture models, transwell and mixed cocultures, were proposed for 7 days with variable HAMSCs: HBMSCs ratios. Then, supernatant from each coculture was used to reverse the deficiency of HBMSCs and human umbilical vein endothelial cells (HUVECs) impaired by high glucose and (GP). We found that glucolipotoxicity caused by GP remarkably inhibited cell proliferation, osteogenic differentiation and superoxide dismutase (SOD) activity, as well as induced the reactive oxygen species (ROS) level in HBMSCs. Meanwhile, glucolipotoxicity suppressed cell proliferation, tube formation capacity and angiogenic potential of HUVECs. Though, HAMSCs/HBMSCs coculture system reduced HBMSCs dysfunction by antioxidant properties and promoted angiogenesis in HUVECs. The mixed HAMSCs/HBMSCs coculture at the optimal ratio of 3/1 showed significantly greater cell proliferation, antioxidant properties, osteogenic and angiogenic differentiation than HBMSCs or HUVECs alone. In conclusion, the current coculture system of HAMSCs/HBMSCs can be a potential therapeutic material for advancing bone and vascular regeneration against DM-induced glucolipotoxicity.Copyright © 2018 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Keyword: diabetes

[Laurine fatty acids, medium fatty acids and triglycerides, hyperlipidemia, resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I , insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral fat cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the metabolic transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: diabetes

BRICHOS domain of Bri2 inhibits islet amyloid polypeptide (IAPP) fibril formation and toxicity in human beta cells.

Aggregation of islet amyloid polypeptide (IAPP) into amyloid fibrils in islets of Langerhans is associated with type 2 , and formation of toxic IAPP species is believed to contribute to the loss of insulin-producing beta cells. The BRICHOS domain of integral membrane protein 2B (Bri2), a transmembrane protein expressed in several peripheral tissues and in the brain, has recently been shown to prevent fibril formation and toxicity of Aβ42, an amyloid-forming peptide in Alzheimer disease. In this study, we demonstrate expression of Bri2 in human islets and in the human beta-cell line EndoC-βH1. Bri2 colocalizes with IAPP intracellularly and is present in amyloid deposits in patients with type 2 . The BRICHOS domain of Bri2 effectively inhibits fibril formation in vitro and instead redirects IAPP into formation of amorphous aggregates. Reduction of endogenous Bri2 in EndoC-βH1 cells with siRNA increases sensitivity to metabolic stress leading to cell death while a concomitant overexpression of Bri2 BRICHOS is protective. Also, coexpression of IAPP and Bri2 BRICHOS in lateral ventral neurons of results in an increased cell survival. IAPP is considered to be the most amyloidogenic peptide known, and described findings identify Bri2, or in particular its BRICHOS domain, as an important potential endogenous inhibitor of IAPP aggregation and toxicity, with the potential to be a possible target for the treatment of type 2 .Copyright © 2018 the Author(s). Published by PNAS.

Keyword: diabetes

Human amnion-derived mesenchymal stem cells promote osteogenesis of human bone marrow mesenchymal stem cells against glucolipotoxicity.

Epidemiological evidence suggests that (DM) is an important factor in promoting periodontitis. It not only affects the attachment of connective tissue but also causes loss of alveolar bone. Hence, there is an urgent need to find an effective treatment for DM-induced bone deficiency. This study aimed to investigate the effects of human amniotic mesenchymal stem cells (HAMSCs) on the proliferation and osteogenic differentiation of DM-induced human bone marrow mesenchymal stem cells (HBMSCs). High glucose and (GP) were used to mimic DM-induced glucolipotoxicity. The proliferation levels were measured using flow cytometry. Alkaline phosphatase activity substrate assays, Alizarin red S staining, and western blotting were used to investigate osteogenic differentiation. Oxidative stress was measured by assaying the levels of reactive oxygen species. This study found that glucolipotoxicity caused by GP remarkably inhibited cell proliferation and osteogenesis, and upregulated the oxidative stress level in HBMSCs. However, HAMSCs attenuated HBMSC dysfunction through antioxidant activity by influencing p38 mitogen-activated protein kinase and vascular endothelial growth factor secretion. In conclusion, our findings indicate that HAMSCs might be suitable for treating DM-mediated bone deficiency.

Keyword: diabetes

Reduces the Autophagic Flux and Insulin Sensitivity Through the Activation of the Free Fatty Receptor 1 (FFAR1) in the Hypothalamic Neuronal Cell Line N43/5.

Chronic consumption of high fat diets (HFDs), rich in saturated fatty acids (SatFAs) like (PA), is associated with the development of obesity and obesity-related metabolic diseases such as type II (T2DM). Previous studies indicate that PA accumulates in the hypothalamus following consumption of HFDs; in addition, HFDs consumption inhibits autophagy and reduces insulin sensitivity. Whether malfunction of autophagy specifically in hypothalamic neurons decreases insulin sensitivity remains unknown. PA does activate the Free Fatty Receptor 1 (FFAR1), also known as G protein-coupled receptor 40 (GPR40); however, whether FFAR1 mediates the effects of PA on hypothalamic autophagy and insulin sensitivity has not been shown. Here, we demonstrate that exposure to PA inhibits the autophagic flux and reduces insulin sensitivity in a cellular model of hypothalamic neurons (N43/5 cells). Furthermore, we show that inhibition of autophagy and the autophagic flux reduces insulin sensitivity in hypothalamic neuronal cells. Interestingly, the inhibition of the autophagic flux, and the reduction in insulin sensitivity are prevented by pharmacological inhibition of FFAR1. Our findings show that dysregulation of autophagy reduces insulin sensitivity in hypothalamic neuronal cells. In addition, our data suggest FFAR1 mediates the ability of PA to inhibit autophagic flux and reduce insulin sensitivity in hypothalamic neuronal cells. These results reveal a novel cellular mechanism linking PA-rich diets to decreased insulin sensitivity in the hypothalamus and suggest that hypothalamic autophagy might represent a target for future T2DM therapies.

Keyword: diabetes

Activator protein-1 and caspase 8 mediate p38α MAPK-dependent cardiomyocyte apoptosis induced by .

Lipoapoptosis of cardiomyocytes may underlie diabetic cardiomyopathy. Numerous forms of cardiomyopathies share a common end-pathway in which apoptotic loss of cardiomyocytes is mediated by p38α mitogen activated protein kinase (MAPK). Although we have previously shown that (PA), a saturated fatty (SFA) elevated in plasma of type 2 and morbid obesity, induces apoptosis in cardiomyocytes via p38α MAPK-dependent signaling, the downstream cascade events that cause cell death remain unknown. The objective of this study\xa0was to investigate mechanisms involved in -induced cardiomyocyte apoptosis. Human adult ventricular cardiomyocyte line (AC16 cells) exposed to high physiological levels of PA for 16\xa0h showed enhanced transcription and phosphorylation of c-fos and c-jun subunits of AP-1 and transcription of caspase 8. When AC16 cells were transfected with small interfering RNA specific against p38α MAPK (si-p38α) for 24 or 48\xa0h, the amplified phosphorylation of c-fos was dose-dependently attenuated, and procaspase 8 was dose-dependently reduced. With translational knockdown of c-fos, PA-induced apoptosis was diminished. Inhibition of caspase 8 for 24\xa0h reduced apoptosis in PA-treated cardiomyocytes. These findings provide evidence for induction of apoptosis in cardiomyocytes exposed to high SFA by a novel pathway requiring activation of c-fos/AP-1 and caspase 8. These results demonstrate how elevated plasma SFA may lead to continual and cumulative loss of cardiomyocytes and potentially contribute to the\xa0development of diabetic cardiomyopathy.

Keyword: diabetes

Fibroblast Growth Factor 21 Stimulates Pancreatic Islet Autophagy via Inhibition of AMPK-mTOR Signaling.

Islet autophagy plays a role in glucose/lipid metabolism in type 2 . Meanwhile, fibroblast growth factor 21 (FGF21) has been found to regulate insulin sensitivity and glucose homeostasis. Whether FGF21 induces islet autophagy, remains to be elucidated. This study aimed to explore the physiological roles and signaling pathways involved in FGF21-stimulated islet autophagy under glucolipotoxic conditions.C57/BL6J mice were fed a standard diet or high-fat diet (HFD) for 12 weeks, and islets were isolated from normal and knockout (KO) mice. Isolated islets and INS-1E cells were exposed to normal and high-concentration glucose and with/without FGF21 or AMPK inhibitor compound C. Real-time PCR, Western blot and immunohistochemistry/transmission electron microscopy were performed for the expression of targeted genes/proteins.HFD-treated mice showed increases in fasting plasma glucose, body weight and impaired glucose tolerance; islet protein expression of FGF21 was induced after HFD treatment. Protein expression levels of FGF21 and LC3-II (autophagy marker) were induced in mouse islets treated with high concentrations of and glucose, while phosphorylation of AMPK was reduced, compared with controls. In addition, induction of LC3-II protein expression was reduced in islets isolated from KO mice. Furthermore, exogenous administration of FGF21 diminished phosphorylation of AMPK and stimulated protein expression of LC3-II. Consistently, compound C significantly induced increased expression of LC3-II protein.Our data indicate that glucolipotoxicity-induced FGF21 activation mediates islet autophagy via AMPK inhibition, and further consolidate the evidence for the FGF21/analog being a pharmacotherapeutic target for obesity and its related T2DM.

Keyword: diabetes

Myristic specifically stabilizes diacylglycerol kinase δ protein in C2C12 skeletal muscle cells.

Decreased levels of the δ isozyme of diacylglycerol kinase (DGK) in skeletal muscle attenuate glucose uptake and, consequently, are critical for the pathogenesis of type 2 . We recently found that free myristic (14:0), but not free (16:0), increased the DGKδ protein levels and enhanced glucose uptake in C2C12 myotube cells. However, it has been unclear how myristic regulates the level of DGKδ2 protein. In the present study, we characterized the myristic -dependent increase of DGKδ protein. A cycloheximide chase assay demonstrated that myristic , but not , markedly stabilized DGKδ protein. Moreover, other DGK isozymes, DGKη and ζ, as well as glucose uptake-related proteins, such as protein kinase C (PKC) α, PKCζ, Akt and glycogen synthase kinase 3β, failed to be stabilized by myristic . Furthermore, DGKδ was not stabilized in cultured hepatocellular carcinoma cells, pancreas carcinoma cells or neuroblastoma cells, and only a moderate stabilizing effect was observed in embryonic kidney cells. A proteasome inhibitor and a lysosome inhibitor, MG132 and chloroquine, respectively, partly inhibited DGKδ degradation, suggesting that myristic prevents, at least in part, the degradation of DGKδ by the ubiquitin-proteasome system and the autophagy-lysosome pathway. Overall, these results strongly suggest that myristic attenuates DGKδ protein degradation in skeletal muscle cells and that this attenuation is fatty -, protein- and cell line-specific. These new findings provide novel insights into the molecular mechanisms of the pathogenesis of type 2 .Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: diabetes

METRNL attenuates lipid-induced inflammation and insulin resistance via AMPK or PPARδ-dependent pathways in skeletal muscle of mice.

Physical activity has many beneficial effects on metabolic disorders, such as obesity, insulin resistance, and . Meteorin-like protein (METRNL), a novel secreted protein homologous to the neurotrophin Metrn, is induced after exercise in the skeletal muscle. Herein, we investigated the effects of METRNL on lipid-mediated inflammation and insulin resistance in skeletal muscle via AMP-activated protein kinase (AMPK) or peroxisome proliferator-activated receptor δ (PPARδ). Treatment with METRNL suppressed inflammatory markers, such as nuclear factor κB (NFκB) nuclear translocation, inhibitory κBα (IκBα) phosphorylation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα and MCP-1). METRNL treatment also attenuated the impaired insulin response both in palmitate-treated differentiated C2C12 cells and the skeletal muscle of high-fat diet (HFD)-fed mice. Furthermore, METRNL administration rescued glucose intolerance and reduced HFD-induced body weight gain in mice; however, METRNL did not affect calorie intake. METRNL treatment increased AMPK phosphorylation and PPARδ expression both in differentiated C2C12 cells and mouse skeletal muscle. siRNA-mediated suppression of AMPK and PPARδ abrogated the suppressive effects of METRNL on palmitate-induced inflammation and insulin resistance. Moreover, METRNL augmented the mRNA expression of fatty oxidation-associated genes, such as carnitine palmitoyltransferase 1 (CPT1), acyl-CoA oxidase (ACO), and fatty binding protein 3 (FABP3). siRNAs for AMPK and PPARδ reversed these changes. In the current study, we report for the first time that METRNL alleviates inflammation and insulin resistance and induces fatty oxidation through AMPK or PPARδ-dependent signaling in skeletal muscle.

Keyword: diabetes

Fatty acids, epigenetic mechanisms and chronic diseases: a systematic review.

Chronic illnesses like obesity, type 2 (T2D) and cardiovascular diseases, are worldwide major causes of morbidity and mortality. These pathological conditions involve interactions between environmental, genetic, and epigenetic factors. Recent advances in nutriepigenomics are contributing to clarify the role of some nutritional factors, including dietary fatty acids in gene expression regulation. This systematic review assesses currently available information concerning the role of the different fatty acids on epigenetic mechanisms that affect the development of chronic diseases or induce protective effects on metabolic alterations.A targeted search was conducted in the PubMed/Medline databases using the keywords "fatty acids and epigenetic". The data were analyzed according to the PRISMA-P guidelines.Consumption fatty acids like n-3 PUFA: EPA and DHA, and MUFA: oleic and palmitoleic was associated with an improvement of metabolic alterations. On the other hand, fatty acids that have been associated with the presence or development of obesity, T2D, pro-inflammatory profile, atherosclerosis and IR were n-6 PUFA, saturated fatty acids (stearic and ), and trans fatty acids (elaidic), have been also linked with epigenetic changes.Fatty acids can regulate gene expression by modifying epigenetic mechanisms and consequently result in positive or negative impacts on metabolic outcomes.

Keyword: diabetes

Nrf2-Mediated Antioxidant Defense and Peroxiredoxin 6 Are Linked to Biosynthesis of Ester of 9-Hydroxystearic .

Fatty esters of hydroxy fatty acids (FAHFAs) are lipid mediators with promising antidiabetic and anti-inflammatory properties that are formed in white adipose tissue (WAT) via de novo lipogenesis, but their biosynthetic enzymes are unknown. Using a combination of lipidomics in WAT, quantitative trait locus mapping, and correlation analyses in rat BXH/HXB recombinant inbred strains, as well as response to oxidative stress in murine models, we elucidated the potential pathway of biosynthesis of several FAHFAs. Comprehensive analysis of WAT samples identified ∼160 regioisomers, documenting the complexity of this lipid class. The linkage analysis highlighted several members of the nuclear factor, erythroid 2 like 2 ()-mediated antioxidant defense system (), lipid-handling proteins (), and the family of flavin containing monooxygenases () as the positional candidate genes. Transgenic expression of and deletion of genes resulted in reduction of ester of 9-hydroxystearic (9-PAHSA) and 11-PAHSA levels, while oxidative stress induced by an inhibitor of glutathione synthesis increased PAHSA levels nonspecifically. Our results indicate that the synthesis of FAHFAs via carbohydrate-responsive element-binding protein-driven de novo lipogenesis depends on the adaptive antioxidant system and suggest that FAHFAs may link activity of this system with insulin sensitivity in peripheral tissues.© 2018 by the American Association.

Keyword: diabetes

Multiplex Surface Plasmon Resonance Imaging-Based Biosensor for Human Pancreatic Islets Hormones Quantification.

arises from secretory defects in vascularized micro-organs known as the islets of Langerhans. Recent studies indicated that furthering our understanding of the paracrine effect of somatostatin on glucose-induced insulin secretion could represent a novel therapeutic avenue for . While many research groups are interested in insulin and glucagon secretion, few are particularly focused on studying the paracrine interaction in islets\' cells, and none on monitoring a secretory fingerprint that contemplates more than two hormones. Surface plasmon resonance imaging can achieve high-throughput and multiplexed biomolecule quantification, making it an ideal candidate for detection of multiple islet\'s secretion products if arrays of hormones can be properly implemented on the sensing surface. In this study, we introduced a multiplex surface plasmon resonance imaging-based biosensor for simultaneous quantification of insulin, glucagon, and somatostatin. Performing this multiplex biosensing of hormones was mainly the result of the design of an antifouling sensing surface comprised by a mixed self-assembly monolayer of CHO-PEG-SH and 16-mercaptohexadecanoic , which allowed it to operate in a complex matrix such as an islet secretome. The limit of detection in multiplex mode was 1 nM for insulin, 4 nM for glucagon, and 246 nM for somatostatin with a total analysis time of 21 min per point, making our approach the first reporting a label-free and multiplex measurement of such a combination of human hormones. This biosensor holds the promise of providing us with a mean for the further understanding of the paracrine effect of somatostatin on glucose-induced insulin secretion and consequently help develop novel therapeutic agents for .

Keyword: diabetes

SAD-A, a downstream mediator of GLP-1 signaling, promotes the phosphorylation of Bad S155 to regulate in\xa0vitro β-cell functions.

The incretin hormone GLP-1 reduces β-cell failure in patients with type 2 . Previous studies demonstrated that GLP-1 activates SAD-A, a member of the AMPK family, to regulate glucose-stimulated secretion (GSIS), but the underlying mechanisms of SAD-A regulation of β-cell functions remain poorly understood. Here, we propose that activation of SAD-A by GLP-1 promotes the phosphorylation of Bad S155, which in turn positively affects GSIS and β-cell survival. Bad therefore appears to be a downstream molecule of a SAD-A pathway that mediates the GLP-1-triggered reduction in β-cell failure. Knockdown of endogenous SAD-A expression significantly exacerbated in\xa0vitro β-cell dysfunction under lipotoxic conditions and promoted lipotoxicity-induced apoptosis, whereas overexpression of SAD-A inhibited β-cell apoptosis. SAD-A silencing increased ER stress and inhibited the autophagic flux, which contributed to β-cell apoptosis. Thus, SAD-A appears to function as a downstream molecule of GLP-1 signaling that results in Bad S155 phosphorylation. This phosphorylation might therefore be involved in the GLP-1-linked protection against β-cell dysfunction and apoptosis.Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

Glucagon-like peptide-1 ameliorates cardiac lipotoxicity in diabetic cardiomyopathy via the PPARα pathway.

Lipotoxicity cardiomyopathy is the result of excessive accumulation and oxidation of toxic lipids in the heart. It is a major threat to patients with . Glucagon-like peptide-1 (GLP-1) has aroused considerable interest as a novel therapeutic target for because it stimulates insulin secretion. Here, we investigated the effects and mechanisms of the GLP-1 analog exendin-4 and the dipeptidyl peptidase-4 inhibitor saxagliptin on cardiac lipid metabolism in diabetic mice (DM). The increased myocardial lipid accumulation, oxidative stress, apoptosis, and cardiac remodeling and dysfunction induced in DM by low streptozotocin doses and high-fat diets were significantly reversed by exendin-4 and saxagliptin treatments for 8\xa0weeks. We found that exendin-4 inhibited abnormal activation of the (PPARα)-CD36 pathway by stimulating protein kinase A (PKA) but suppressing the Rho-associated protein kinase (ROCK) pathway in DM hearts, (PA)-treated rat h9c2 cardiomyocytes (CMs), and isolated adult mouse CMs. Cardioprotection in DM mediated by exendin-4 was abolished by combination therapy with the PPARα agonist wy-14643 but mimicked by PPARα gene deficiency. Therefore, the PPARα pathway accounted for the effects of exendin-4. This conclusion was confirmed in cardiac-restricted overexpression of PPARα mediated by adeno-associated virus serotype-9 containing a cardiac troponin T promoter. Our results provide the first direct evidence that GLP-1 protects cardiac function by inhibiting the ROCK/PPARα pathway, thereby ameliorating lipotoxicity in diabetic cardiomyopathy.© 2018 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.

Keyword: diabetes

Saturated fatty acids bound to albumin enhance osteopontin expression and cleavage in renal proximal tubular cells.

Osteopontin (OPN) is one of the proinflammatory cytokines upregulated in the kidneys of diabetic animals and patients with nephropathy. An increase in urinary albumin and albumin-bound fatty acids (FA) presents a proinflammatory environment to the proximal tubules in proteinuric kidney diseases including diabetic nephropathy. This study was designed to examine if FA overload could stimulate OPN expression and cleavage in renal tubule epithelial cells. OPN gene and protein expression was examined in the kidney of Zucker diabetic (ZD) rats and cultured proximal tubular cells exposed to either bovine serum albumin (BSA) or BSA conjugated with (PA), the most abundant saturated plasma FA. Real-time PCR analysis confirmed an upregulation of renal cortical OPN gene correlated with albuminuria and nephropathy progression in ZD rats at the age of 7-20 weeks. Immunofluorescence staining of kidney sections revealed a massive induction of OPN protein in albumin-overloaded proximal tubules of ZD rats. A significant increase in both intact and cleaved OPN proteins was further demonstrated in the diabetic kidney and urine samples, which was attenuated by antiproteinuric treatment with losartan, an angiotensin II receptor blocker. When exposed to fatty -free BSA, NRK-52E cells exhibited an increase in protein levels of full-length and cleaved OPN. Moreover, the increase in OPN fragments was greatly enhanced in the presence of PA (250-500 µM). Together, our results support a stimulatory effect of albumin and conjugated FA on OPN expression and cleavage in renal tubule epithelial cells. Thus, besides lowering albuminuria/proteinuria, mitigating circulating FAs may be an effective intervention for preventing and slowing down the progression of nephropathy associated with obesity and type 2 .

Keyword: diabetes

Palmitate induces nitric oxide production and inflammatory cytokine expression in zebrafish.

Inflammation markers in zebrafish embryos reflect a toxic response that is common to other animal models and humans. Free fatty acids (FFAs) are known to cause damage in various tissues by inducing inflammation. In this study, we investigated whether a FFA (palmitate) induces inflammation in zebrafish embryos. Nitrous oxide (NO) production and cyclooxygenase-2 (COX-2) mRNA expression were increased in palmitate-treated zebrafish embryos in a dose-dependent manner. mRNA expression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF- α), were also increased. Additionally, the mRNA expression of p65 nuclear factor-kB and I-kB-α were significantly increased after palmitate-treatment. Increased reactive oxygen species (ROS) expression was observed in palmitate-treated zebrafish embryos as well as pericardial edema. Additionally, mRNA expression of pro-inflammatory cytokines were increased in zebrafish liver and pancreas fed with palmitate-contained diet. Taken together, these results indicated that palmitate increases pro-inflammatory mediators in zebrafish embryos, suggesting that zebrafish could be an alternative animal model for inflammatory disease including .Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: diabetes

Resveratrol reduces intracellular reactive oxygen species levels by inducing autophagy through the AMPK-mTOR pathway.

Oxidative stress induced by free fatty aggravates endothelial injury, which leads to diabetic cardiovascular complications. Reduction of intracellular oxidative stress may attenuate these pathogenic processes. The dietary polyphenol resveratrol reportedly exerts potential protective effects against endothelial injury. This study determined whether resveratrol can reduce the (PA)-induced generation of reactive oxygen species (ROS) and further explored the underlying molecular mechanisms. We found that resveratrol significantly reduced the PA-induced endothelial ROS levels in human aortic endothelial cells. Resveratrol also induced endothelial cell autophagy, which mediated the effect of resveratrol on ROS reduction. Resveratrol stimulated autophagy via the AMP-activated protein kinase (AMPK)-mTOR pathway. Taken together, these data suggest that resveratrol prevents PA-induced intracellular ROS by autophagy regulation via the AMPK-mTOR pathway. Thus, the induction of autophagy by resveratrol may provide a novel therapeutic candidate for cardioprotection in metabolic syndrome.

Keyword: diabetes

Celastrol Reverses -Induced Insulin Resistance in HepG2 Cells via Restoring the miR-223 and GLUT4 Pathway.

The natural triterpenoid compound celastrol ameliorates insulin resistance (IR) in animal models, but the underlying molecular mechanism is unclear. In this study, we investigated how celastrol regulates IR.The HepG2 cellular IR model was initially established with (PA). The expression and activity of glucose transporter 4 (GLUT4), insulin receptor substrate-1 (IRS1) and 9 microRNAs (miRNAs) (miR-7, -34a, -96, -113, -126, -145, -150, -223 and -370) were detected before and after celastrol treatment using the PA-induced HepG2 IR model.The results showed that 250\u2009µM PA for ≥2 days was optimal for inducing IR in HepG2 cells; 600\u2009nM celastrol significantly attenuated the PA-induced IR in HepG2 cells. The PA-induced GLUT4 and IRS1 downregulation and Ser307 phosphorylation on IRS1 was reversed by subsequent treatment with 600\u2009nM celastrol for 6\u2009h. We next investigated which IR-related miRNAs were possible upstream regulators of celastrol-mediated reversal of PA-induced HepG2 IR. Two miRNAs, miR-150 and -223, were significantly downregulated by PA and were re-raised by subsequent celastrol treatment; and miR-223 was upstream of miR-150. Moreover, knocking down miR-223 abolished celastrol\'s anti-IR effects in the PA-induced model.Collectively, our results demonstrated that celastrol reverses PA-induced IR-related alterations, in part via miR-223 in HepG2 cells. Further investigation is warranted for establishing the clinical potential of celastrol in treating IR-related disorders.Copyright © 2019. Published by Elsevier Inc.

Keyword: diabetes

Dietary modulation of energy homoeostasis and metabolic-inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-inflammation, within the context of obesity, type-2 (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular energy metabolism is a key determinant of inflammation. Whilst metabolic-inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: diabetes

TAp63 is correlated with chronic inflammation in patients with newly diagnosed type 2 .

To investigate TAp63 expression in patients with type 2 (T2DM) and the potential correlations between TAp63 and proinflammatory cytokines production and other clinical parameters.Peripheral blood mononuclear cells (PBMCs) and plasma were collected from 72 T2DM (cases) and 72 healthy subjects (controls). Fasting blood glucose (FBG), fasting insulin (FIN) and a blood lipid profile were measured. The homeostasis model assessment (HOMA) was used to estimate insulin resistance (IR). Plasma tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were determined. PBMCs isolated from healthy subjects were cultured with or without 33.3 mmol/l glucose or 0.5 mmol/l (PA) for 6 h, 24 h, 48 h, and 72 h. The expression of TAp63 at mRNA and protein levels in PBMCs was analyzed using real-time qRT-PCR and western blots, respectively.TAp63 expression was significantly lower in T2DM patients compared with that of the controls. In addition, TAp63 expression showed a negative correlation with FBG, FIN, HbA1c, HOMA-IR, FFAs, TNF-α, and IL-6 levels. Treatment with 33.3 mmol/l glucose or 0.5 mmol/l PA increased TAp63 expression in the cultured PBMCs.TAp63 level may be correlated with chronic inflammatory state and perturbed glucose and lipid metabolism in T2DM.Copyright © 2018. Published by Elsevier Inc.

Keyword: diabetes

Hydroxystearic Acids Activate GPR40, Which Is Involved in Their Beneficial Effects on Glucose Homeostasis.

hydroxystearic acids (PAHSAs) are endogenous lipids with anti-diabetic and anti-inflammatory effects. PAHSA levels are reduced in serum and adipose tissue of insulin-resistant people and high-fat diet (HFD)-fed mice. Here, we investigated whether chronic PAHSA treatment enhances insulin sensitivity and which receptors mediate PAHSA effects. Chronic PAHSA administration in chow- and HFD-fed mice raises serum and tissue PAHSA levels ∼1.4- to 3-fold. This improves insulin sensitivity and glucose tolerance without altering body weight. PAHSA administration in chow-fed, but not HFD-fed, mice augments insulin and glucagon-like peptide (GLP-1) secretion. PAHSAs are selective agonists for GPR40, increasing Ca flux, but not intracellular cyclic AMP. Blocking GPR40 reverses improvements in glucose tolerance and insulin sensitivity in PAHSA-treated chow- and HFD-fed mice and directly inhibits PAHSA augmentation of glucose-stimulated insulin secretion in human islets. In contrast, GLP-1 receptor blockade in PAHSA-treated chow-fed mice reduces PAHSA effects on glucose tolerance, but not on insulin sensitivity. Thus, PAHSAs activate GPR40, which is involved in their beneficial metabolic effects.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Resveratrol affects hepatic gluconeogenesis via histone deacetylase 4.

The aim of this study was to determine whether resveratrol (Rev) affects the expression, phosphorylation, and nuclear and cytoplasmic distribution of histone deacetylase 4 (HDAC4), which in turn affects gluconeogenesis in hepatocytes under an insulin-resistant state.HepG2 cells were treated with 0.25 mmol/L (PA) to establish an insulin resistance model. The cells were divided into five groups: control, PA, PA + Rev 100 µM, PA + Rev 50 µM, and PA + Rev 20 µM. After treatment for 24 hours, mRNA and protein expression levels of gluconeogenesis pathway-related molecules and HDAC4 were examined. Next, HepG2 cells were transfected with siRNA-HDAC4. The cells were divided into control, PA, PA + Rev 20 µM, PA + Rev 20 µM +siRNA-HDAC4 negative control, and PA + Rev 20 µM +siRNA-HDAC4 knockdown groups to determine the expression of gluconeogenesis pathway proteins.Compared with the control group, the gluconeogenesis pathway-related molecules, glucose-6-phosphatase catalytic subunit (G6PC), phosphoenolpyruvate carboxykinase 1 (PCK1) and forkhead box protein O1 (FOXO1), were increased, and the phosphorylation of FOXO1 decreased after PA treatment. The p-HDAC4 level decreased with the increase in HDAC4 in the nucleus and the decrease in HDAC4 in the cytoplasm in the PA group. Treatment with Rev 20 µM suppressed gluconeogenesis and promoted HDAC4 shuttling into the cytoplasm from the nucleus. However, 100 and Rev 50 µM exerted the opposite effects. Finally, after HDAC4 knockdown, the expression levels of the key gluconeogenesis molecules, G6PC, PCK1, and FOXO1, were increased, and p-FOXO1 was decreased, indicating that gluconeogenesis was enhanced.A low concentration of Rev inhibited gluconeogenesis under insulin-resistance conditions via translocation of HDAC4 from the nucleus to the cytoplasm.

Keyword: diabetes

Adipose tissue dysfunction is associated with low levels of the novel Hydroxystearic Acids.

Adipose tissue dysfunction is considered an important contributor to systemic insulin resistance and Type 2 (T2D). Recently, a novel family of endogenous lipids, hydroxy stearic acids (PAHSAs), was discovered. These have anti-diabetic and anti-inflammatory effects in mice and are reduced in serum and adipose tissue of insulin resistant humans. In the present study, we investigate if adipose tissue dysfunction is associated with reduced PAHSA levels in human subjects and if PAHSAs influence adipocyte differentiation. Our results show that low expression of adipocyte GLUT4 and adipocyte hypertrophy, markers of adipose tissue dysfunction, are associated with reduced expression of key enzymes for de novo lipogenesis and adipose tissue levels of PAHSAs in human subjects. We also show that GLUT4 is not only a marker of adipose tissue dysfunction, but may be causally related to the observed impairments. PAHSAs may also act locally in the adipose tissue to improve adipogenesis through a mechanism bypassing direct activation of peroxisome proliferator-activated receptor (PPARγ). The discovery of PAHSAs and our current results provide novel insights into positive effects of lipid species in adipose tissue and mechanisms by which dysfunctional adipose tissue is associated with insulin resistance and risk of developing T2D.

Keyword: diabetes

Mitochondrial dysfunction and inhibition of myoblast differentiation in mice with high-fat-diet-induced pre-.

Pre- is characterized by impaired glucose tolerance (IGT) and/or impaired fasting glucose. Impairment of skeletal muscle function is closely associated with the progression of . However, the entire pathological characteristics and mechanisms of pre- in skeletal muscle remain fully unknown. Here, we established a mouse model of pre-, in which 6-week-old male C57BL6/J mice were fed either normal diet or high-fat diet (HFD) for 8 or 16 weeks. Both non-fasting and fasting glucose levels and the results of glucose and insulin tolerance tests showed that mice fed an 8-week HFD developed pre- with IGT; whereas mice fed a 16-week HFD presented with impaired fasting glucose and impaired glucose tolerance (IFG-IGT). Mice at both stages of pre- displayed decreased numbers of mitochondria in skeletal muscle. Moreover, IFG-IGT mice exhibited decreased mitochondrial membrane potential and ATP production in skeletal muscle and muscle degeneration characterized by a shift in muscle fibers from predominantly oxidative type I to glycolytic type II. Western blotting and histological analysis confirmed that myoblast differentiation was only inhibited in IFG-IGT mice. For primary skeletal muscle satellite cells, inhibition of differentiation was observed in -induced insulin resistance model. Moreover, enhanced myoblast differentiation increased glucose uptake and insulin sensitivity. These findings indicate that pre- result in mitochondrial dysfunction and inhibition of myoblast differentiation in skeletal muscle. Therefore, interventions that enhance myoblast differentiation may improve insulin resistance of at the earlier stage.© 2018 Wiley Periodicals, Inc.

Keyword: diabetes

Clinopodium chinense Attenuates -Induced Vascular Endothelial Inflammation and Insulin Resistance through TLR4-Mediated NF- B and MAPK Pathways.

Elevated (PA) levels are associated with the development of inflammation, insulin resistance (IR) and endothelial dysfunction. Clinopodium chinense (Benth.) O. Kuntze has been shown to lower blood glucose and attenuate high glucose-induced vascular endothelial cells injury. In the present study we investigated the effects of ethyl acetate extract of C. chinense (CCE) on PA-induced inflammation and IR in the vascular endothelium and its molecular mechanism. We found that CCE significantly inhibited PA-induced toll-like receptor 4 (TLR4) expression in human umbilical vein endothelial cells (HUVECs). Consequently, this led to the inhibition of the following downstream adapted proteins myeloid differentiation primary response gene 88, Toll/interleukin-1 receptor domain-containing adaptor-inducing interferon- and TNF receptor-associated factor 6. Moreover, CCE inhibited the phosphorylation of Ikappa B kinase , nuclear factor kappa-B (NF- B), c-Jun N-terminal kinase, extracellular regulated protein kinases, p38-mitogen-activated protein kinase (MAPK) and subsequently suppressed the release of tumor necrosis factor- , interleukin-1 (IL-1 ) and IL-6. CCE also inhibited IRS-1 serine phosphorylation and ameliorated insulin-mediated tyrosine phosphorylation of IRS-1. Moreover, CCE restored serine/threonine kinase and endothelial nitric oxide synthase (eNOS) activation and thus increased insulin-mediated nitric oxide (NO) production in PA-treated HUVECs. This led to reverse insulin mediated endothelium-dependent relaxation, eNOS phosphorylation and NO production in PA-treated rat thoracic aortas. These results suggest that CCE can significantly inhibit the inflammatory response and alleviate impaired insulin signaling in the vascular endothelium by suppressing TLR4-mediated NF- B and MAPK pathways. Therefore, CCE can be considered as a potential therapeutic candidate for endothelial dysfunction associated with IR and .

Keyword: diabetes

The Dynamic Effects of Isosteviol on Insulin Secretion and Its Inability to Counteract the Impaired β-Cell Function during Gluco-, Lipo-, and Aminoacidotoxicity: Studies In Vitro.

Isosteviol (ISV), a diterpene molecule, is an isomer of the backbone structure of a group of substances with proven antidiabetic capabilities. The aim of this study was to investigate if ISV elicits dynamic insulin release from pancreatic islets and concomitantly is able to ameliorate gluco-, lipo-, and aminoacidotoxicity in clonal β-cell line (INS-1E) in relation to cell viability and insulin secretion. Isolated mice islets placed into perifusion chambers were perifused with 3.3 mM and 16.7 mM glucose with/without 10 M ISV. INS-1E cells were incubated for 72 h with either 30 mM glucose, 1 mM palmitate or 10 mM leucine with or without 10 M ISV. Cell viability was evaluated with a Cytotoxic Fluoro-test and insulin secretion was measured in Krebs-Ringer Buffer at 3.3 mM and 16.7 mM glucose. In the presence of 3.3 mM glucose, 10 M ISV did not change basal insulin secretion from perifused islets. However, at a high glucose level of 16.7 mM, 10 M ISV elicited a 2.5-fold increase (-ISV: 109.92 ± 18.64 ng/mL vs. +ISV: 280.15 ± 34.97 ng/mL; < 0.01). After 72 h gluco-, lipo-, or aminoacidotoxicity in INS-1E cells, ISV treatment did not significantly affect cell viability (glucotoxicity, -ISV: 19.23 ± 0.83%, +ISV: 18.41 ± 0.90%; lipotoxicity, -ISV: 70.46 ± 3.15%, +ISV: 65.38 ± 2.81%; aminoacidotoxicity: -ISV: 8.12 ± 0.63%; +ISV: 7.75 ± 0.38%, all nonsignificant). ISV did not improve impaired insulin secretion (glucotoxicity, -ISV: 52.22 ± 2.90 ng/mL, +ISV: 47.24 ± 3.61 ng/mL; lipotoxicity, -ISV: 19.94 ± 4.10 ng/mL, +ISV: 22.12 ± 3.94 ng/mL; aminoacidotoxicity: -ISV: 32.13 ± 1.00 ng/mL; +ISV: 30.61 ± 1.54 ng/mL, all nonsignificant). In conclusion, ISV acutely stimulates insulin secretion at high but not at low glucose concentrations. However, ISV did not counteract cell viability or cell dysfunction during gluco-, lipo-, or aminoacidotoxicity in INS-1E cells.

Keyword: diabetes

Primary defects in lipid handling and resistance to exercise in myotubes from obese donors with and without type 2 .

Several studies have shown that human primary myotubes retain the metabolic characteristic of their donors in vitro. Others and we have demonstrated a reduced lipid turnover and fat oxidation rate in myotubes derived from obese donors without and with type 2 (T2D). Because exercise is known to increase fat oxidative capacity in skeletal muscle, we investigated if in vitro exercise could restore primary defects in lipid handling in myotubes of obese without and with T2D compared to lean non-diabetic donors. Primary myotubes cultures were derived from biopsies of lean, obese and T2D subjects. One single bout of long-duration exercise was mimicked in vitro by electrical pulse stimulation (EPS) for 24 h. Lipid handling was measured using radiolabeled palmitate, Metabolic gene expression by real-time qPCR, and proteins by western blot. We first show that myotubes from obese and T2D donors had increased uptake and incomplete oxidation of palmitate. This was associated with reduced mitochondrial respiratory chain complex II, III and IV protein expression in myotubes from obese and T2D subjects. EPS stimulated palmitate oxidation in lean donors, while myotubes from obese and T2D donors were refractory to this effect. Interestingly, EPS increased total palmitate uptake in myotubes from lean donors while myotubes from T2D donors had a reduced rate of palmitate uptake into complex lipids and triacylglycerols. Novelty: • Myotubes from obese and T2D donors are characterized by primary defects in handling. • Both obese and T2D myotubes are partially refractory to the beneficial effect of exercise on lipid handling.

Keyword: diabetes

Antagonistic interaction between Nodal and insulin modulates pancreatic β-cell proliferation and survival.

Insulin signaling pathway in β-cell is essential to promote β-cells proliferation and survival, while Nodal-ALK7-Smad3 signaling involves β-cells apoptosis. We attempted to address inter-relationship between Nodal and insulin in modulating β-cell proliferation and apoptosis.Using INS-1 β-cells and isolated rat islets, we examined the effects of Nodal, insulin, or the two combined on β-cell proliferation and/or apoptosis.The β-cells under high-glucose or palmitate conditions showed significant up-regulation of Nodal expression and activation of its downstream signaling pathway resulted in increased cleaved caspase-3. Insulin treatment led to significantly attenuated Nodal-induced cell apoptotic pathway. Similar results were found in directly Nodal-treated β-cell that insulin could partially block Nodal-induced up-regulation of ALK7-Smad3-caspase-3 signaling pathways with significantly attenuated β-cell apoptosis. Interestingly, we found that insulin-induced Akt activation and downstream molecules including GSK-3β, β-catenin and ERK1/2 was significantly attenuated by the co-treatment with Nodal, resulted in decreased cell proliferation. Furthermore, Nodal decreased glucose-evoked calcium influx and played a negative role during glucose-stimulated insulin secretion in the β-cells. Immunocytochemistry studies showed that Nodal treatment translocated Smad3 from cytosol mostly to the nucleus; however, co-treatment with insulin significantly decreased Smad3 nuclear localization. Co-immunoprecipitation experiments showed a directly interaction between Smad3 and Akt, and this interaction was enhanced by co-treatment with insulin.Our data suggest that the antagonistic interaction between Nodal and insulin has a role in the regulation of β-cell mass and secretion.

Keyword: diabetes

Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARα/PPARγ coactivator-1α pathway.

Nuciferine, an alkaloid found in Nelumbo nucifera leaves, alleviates dyslipidemia in vivo. However, whether it improves liver injury in diabetic conditions and the underlying mechanism is unclear. The present study aimed to investigate the effects of nuciferine on lipid and glucose metabolism in a murine model of Type 2 (T2DM) and to determine the underlying mechanisms of these effects.A murine model of T2DM was induced by high-fat diet (HFD) feeding combined with streptozocin (STZ) injections, and the diabetic mice were treated with nuciferine in their food. The underlying mechanism of the anti-steatotic effect of nuciferine was further explored in HepG2 hepatocytes cultured with . Major signalling profiles involved in fatty oxidation were then evaluated, using Western blot, RT-qPCR and si-RNA techniques, along with immunohistochemistry.Nuciferine restored impaired glucose tolerance and insulin resistance in diabetic mice. Hepatic levels of total cholesterol, triglycerides and LDL were decreased, as were the number of lipid droplets, by nuciferine treatment. Furthermore, nuciferine up-regulated β-oxidation related genes in livers of diabetic mice. Luciferase reporter cell assay showed that nuciferine directly reversed -induced inhibition of PPARα transcriptional activity. Silencing PPARγ coactivator-1α (PGC1α) expression in HepG2 cells abolished the effects of nuciferine in accelerating β-oxidation.Nuciferine improved lipid profile and attenuated hepatic steatosis in HFD/STZ-induced diabetic mice by activating the PPARα/PGC1α pathway. Nuciferine may be a potentially important candidate in improving hepatic steatosis and the management of T2DM.© 2018 The British Pharmacological Society.

Keyword: diabetes

Low molecular weight fucoidan attenuates liver injury via SIRT1/AMPK/PGC1α axis in db/db mice.

Non-alcoholic fatty-liver disease (NAFLD), caused by elevated hepatic lipids, inflammation and oxidative stress, is the most common liver disease globally. Low molecular weight fucoidan (LMWF), a sulfated polysaccharide extracted from brown seaweeds, has shown strong anti-inflammatory and antioxidant activities, which has not been explored in -induced NAFLD. Therefore, the present study sought to determine whether LMWF protects obese diabetic db/db mice against NAFLD. Results showed LMWF administration decreased plasma level of alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride, as well as alleviated hepatic accumulation of triglyceride and total cholesterol in db/db mice. LMWF also ameliorated hepatic oxidative stress by suppressing superoxide production and lipid peroxidation, and increasing catalase and superoxide dismutase activity in the liver of db/db mice. Furthermore, LMWF down-regulated several pro-inflammatory cytokines and transcription factor, and up-regulated the anti-inflammatory adiponectin. These changes were accompanied by the activation of hepatic SIRT1/AMPK/PGC1α signaling with LMWF treatment. In addition, blocking SIRT1 or AMPK by inhibitor notably abolished LMWF-elicited protection against -induced oxidative stress and inflammation in hepatocytes. These results suggest LMWF prevents NAFLD in db/db mice by activation of SIRT1/AMPK/PGC1α signaling pathway, which prevents lipotoxicity-related oxidative stress and inflammation. Therefore, LMWF provides a potential supplementary treatment for obesity/-induced NAFLD.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Triterpenoids from Hibiscus sabdariffa L.\xa0with PPARδ/γ Dual Agonist Action: In Vivo, In Vitro and In Silico Studies.

is a medicinal plant consumed as a diuretic and anti-obesity remedy. Several pharmacological studies have shown its beneficial effects in metabolism. Peroxisome proliferator-activated receptors and may play a role in the actions of These nuclear receptors regulate lipid and glucose metabolism and are therapeutic targets for type 2 . This research aimed to perform a phytochemical study guided by a bioassay from to identify compounds with peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor agonist activity, supported by messenger ribonucleic expression, molecular docking, lipid accumulation, and an antihyperglycemic effect. An oral glucose tolerance test in mice with the aqueous extract of and the dichloromethane extract of was performed. The dichloromethane extract of exhibited an antihyperglycemic effect. The dichloromethane extract of was fractioned, and four fractions were evaluated in 3T3-L1 adipocytes on peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 messenger ribonucleic expression. Fraction F3 exhibited peroxisome proliferator-activated receptor / dual agonist activity, and a further fractionation yielded two subfractions, F3-1 and F3-2, which also increased peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor expression. Subfractions were analyzed by GC/MS. The main compounds identified in F3-1 were linoleic , oleic , and , while in F3-2, the main compounds identified were -amyrin and lupeol. These molecules were subjected to molecular docking analysis. -Amyrin and lupeol showed the highest affinity. Moreover, both produced an increase in peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 expression. Additionally, -amyrin and lupeol decreased lipid accumulation in 3T3-L1 adipocytes and blood glucose in mice. Until now, -amyrin and lupeol have not been reported with activity on peroxisome proliferator-activated receptors. This study provides evidence that -amyrin and lupeol possess antidiabetic effects through a peroxisome proliferator-activated receptor / dual agonist action.Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

The effect of enterolactone on sphingolipid pathway and hepatic insulin resistance development in HepG2 cells.

Obesity and type 2 , correlate with increased tissue concentration of sphingolipids, which directly interfere with insulin signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid metabolism and decrease hepatic insulin resistance development in a lipid overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and insulin signaling pathway proteins and glucose metabolism regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of insulin-stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher fatty acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Sphingomyelinase Down-regulation Alleviates Vascular Endothelial Insulin Resistance in Diabetic Rats.

Insulin resistance in endothelial cells contributes to the development of cardiovascular disease in patients with type 2 . sphingomyelinase (ASM) is a soluble glycoprotein which plays a vital role in the development and progression of various diseases such as cardiovascular and metabolic diseases. However, it remains unknown if ASM regulates insulin resistance in vascular endothelial cells in type 2 . ASM down-regulation with gene silencing and selective inhibitor amitriptyline was used in the rat aortic endothelial cells (RAECs) treated with (PA), a common saturated free fatty , which is thought to be the major cause of insulin resistance. It was shown that ASM down-regulation increased glucose uptake and glucose transporter-4 (Glut4) expression and reversed the phosphorylation of pIRS-1-ser307 and AKT-ser473 via ceramide, consequently resulting in the decrease of the production of endothelial nitric oxide synthase (eNOS) and nitric oxide in PA-induced RAECs. We further found that ASM down-regulation blocked the Nox2- and Nox4-dependent superoxide (O ) generation, which regulated glucose metabolism in RAECs during PA stimulation. In vivo, amitriptyline relieved the vasodilatory response to acetylcholine and restored the level of ceramide, Nox2 and Nox4 in the aorta endothelium of high-fat diet-fed rats following an injection of streptozotocin. Taken together, these results suggest that ASM down-regulation can improve endothelial insulin resistance which is attributed to inhibiting redox signalling in RAECs. Thus, these data support the idea that ASM is a promising clinical biomarker and potential therapeutic target for diabetic vascular complication.© 2018 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

Keyword: diabetes

Saturated fatty combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated fatty (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic inflammation, a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Keyword: diabetes

Liraglutide ameliorates palmitate-induced insulin resistance through inhibiting the IRS-1 serine phosphorylation in mouse skeletal muscle cells.

A reduction in insulin-stimulated glucose uptake in skeletal muscles is a characteristic of insulin resistance and type 2 (T2DM). The glucagon-like peptide (GLP)-1 agonist liraglutide can reduce blood glucose levels in individuals with T2DM. However, its effect on insulin-induced glucose metabolism in the skeletal muscle of insulin resistance is unknown. We investigated the effects and action mechanisms of liraglutide on insulin resistance (IR) in the\xa0skeletal muscle cells treatment with (PA).The cell-surface GLUT4myc levels were determined by an antibody-coupled colorimetric assay. The phosphorylation levels of Akt, PI3K(p85α), AS160, IRS1, IKK, and JNK were determined by western blotting. The quantifications of mRNA levels of TNFα, IL-1β, and IL-6 were determined by real-time PCR. Analysis of variance was used for data analysis.PA elevated not only phosphorylation of JNK, IRS1 serines, and IKKα/β, but also the expression of IL-6,\xa0TNFα and IL-1β in C2C12-GLUT4myc cells. PA can reduce phosphorylation of IRS1 tyrosine. These effects of PA were reversed by liraglutide. In addition, liraglutide can reverse PA-decreased insulin-stimulated cell-surface GLUT4 levels, Akt, PI3K(p85α), and AS160 phosphorylation.Liraglutide can enhance insulin-induced GLUT4 translocation by inhibiting IRS1 serine phosphorylation in PA-treated muscle cells.

Keyword: diabetes

Palmitate-Induced Insulin Hypersecretion and Later Secretory Decline Associated with Changes in Protein Expression Patterns in Human Pancreatic Islets.

In obese children with high circulating concentrations of free fatty palmitate, we have observed that insulin levels at fasting and in response to a glucose challenge were several times higher than in obese children with low concentrations of the fatty as well as in lean controls. Declining and even insufficient insulin levels were observed in obese adolescents with high levels of the fatty . In isolated human islets exposed to palmitate we have observed insulin hypersecretion after 2 days exposure. In contrast, insulin secretion from the islets was reduced after 7 days culture in the presence of the fatty . This study aims\xa0at identifying islet-related biological events potentially linked with the observed insulin hypersecretion and later secretory decline in these obese children and adolescents using the islet model. We analyzed protein expression data obtained from human islets exposed to elevated palmitate levels for 2 and 7 days by an improved methodology for statistical analysis of differentially expressed proteins. Protein profiling of islet samples by liquid chromatography-tandem mass spectrometry identified 115 differentially expressed proteins (DEPs). Several DEPs including sorcin were associated with increased glucose-stimulated insulin secretion\xa0in islets after 2 days of exposure to palmitate. Similarly, several metabolic pathways including altered protein degradation, increased autophagy, altered redox condition, and hampered insulin processing were coupled to the functional impairment of islets after 7 days of culture in the presence of palmitate. Such biological events, once validated in the islets, may give rise to novel treatment strategies aiming at normalizing insulin levels in obese children with high palmitate levels, which may reduce or even prevent obesity-related type 2 .

Keyword: diabetes

Hsp74/14-3-3σ Complex Mediates Centrosome Amplification by High Glucose, Insulin, and .

It has been reported recently that type 2 promotes centrosome amplification via 14-3-3σ/ROCK1 complex. In the present study, 14-3-3σ interacting proteins are characterized and their roles in the centrosome amplification by high glucose, insulin, and are investigated. Co-immunoprecipitation in combination with MS analysis identified 134 proteins that interact with 14-3-3σ, which include heat shock 70 kDa protein 4 (Hsp74). Gene ontology analyses reveal that many of them are enriched in binding activity. Kyoto Encyclopedia of Genes and Genomes analysis shows that the top three enriched pathways are ribosome, carbon metabolism, and biosynthesis of amino acids. Molecular and functional investigations show that the high glucose, insulin, and increase the expression and binding of 14-3-3σ and Hsp74 as well as centrosome amplification, all of which are inhibited by knockdown of 14-3-3σ or Hsp74. Moreover, molecular docking analysis shows that the interaction between the 14-3-3σ and the Hsp74 is mainly through hydrophobic contacts and a lesser degree ionic interactions and hydrogen bond by different amino acids residues. In conclusion, the results suggest that the experimental treatment triggers centrosome amplification via upregulations of expression and binding of 14-3-3σ and Hsp74.© 2019 The Authors. Proteomics published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: diabetes

Overexpression of heart-type fatty binding protein enhances fatty -induced podocyte injury.

Deregulated lipid metabolism is a characteristic of metabolic diseases including type 2 and obesity, and likely contributes to podocyte injury and end-stage kidney disease. Heart-type fatty binding protein (H-FABP) was reported to be associated with lipid metabolism. The present study investigated whether H-FABP contributes to podocyte homeostasis. Podocytes were transfected by lentiviral vector to construct a cell line which stably overexpressed H-FABP. Small interfering RNA capable of effectively silencing H-FABP was introduced into podocytes to construct a cell line with H-FABP knockdown. Certain groups were treated with (PA) and the fat metabolism, as well as inflammatory and oxidative stress markers were measured. PA accelerated lipid metabolism derangement, inflammatory reaction and oxidative stress in podocytes. Overexpression of H-FABP enhanced the PA-induced disequilibrium in podocytes. The mRNA and protein expression levels of acyl-coenzyme A oxidase 3 and monocyte chemotactic protein 1, and the protein expression levels of 8-hydroxy-2\'-deoxyguanosine and 4-hydroxynonenal were upregulated in the H-FABP overexpression group, while the mRNA and protein expression of peroxisome proliferator activated receptor α was downregulated. Knockdown of H-FABP inhibited the PA-induced injury and lipid metabolism derangement, as well as the inflammatory reaction and oxidative stress in podocytes. These results indicated that overexpression of H-FABP enhances fatty -induced podocyte injury, while H-FABP inhibition may represent a potential therapeutic strategy for the prevention of lipid metabolism-associated podocyte injury.

Keyword: diabetes

Lipopolysaccharide and synergistically induced MCP-1 production via MAPK-meditated TLR4 signaling pathway in RAW264.7 cells.

Obesity increases the risk of developing . Clinical studies suggest that risk factors like (PA) and lipopolysaccharide (LPS) exist simultaneously in with obesity. Combination of PA and LPS even at low concentration can induce strong inflammatory reaction. Monocyte chemoattractant protein-1 (MCP-1) is an important inflammatory chemokine related to insulin resistance and type II . Our previous study using PCR array revealed that LPS and PA synergistically induce MCP-1 mRNA expression in macrophage cells RAW264.7, while the protein expression of MCP-1 in this case was not investigated. Moreover, the underling mechanism in the synergistic effect of MCP-1 expression or production induced by treatment of LPS and PA combination remains unclear.Protein secretion of MCP-1 was measured by the enzyme-linked immunosorbent assay (ELISA) and mRNA levels of MCP-1 and Toll-like receptor 4 (TLR4) were measured by real-time PCR. Statistical analysis was conducted using SPSS software.LPS could increase MCP-1 transcription as well as secretion in RAW264.7, and PA amplified this effect obviously. Meanwhile, combination of LPS with PA increased TLR4 mRNA expression while LPS alone or PA alone could not, TLR4 knockdown inhibited MCP-1 transcription/secretion induced by LPS plus PA. Moreover, not NF-κB inhibitor but inhibitors of mitogen-activated protein kinase (MAPK) signaling pathways, including c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 MAPK were found to block MCP-1 generation stimulated by LPS plus PA.LPS and PA synergistically induced MCP-1 secretion in RAW264.7 macrophage cells, in which MCP-1 transcription mediated by MAPK/TLR4 signaling pathways was involved. Combined treatment of PA and LPS in RAW264.7 cells mimics the situation of with obesity that has higher level of PA and LPS, MAPK/TLR4/ MCP-1 might be potential therapeutic targets for with obesity.

Keyword: diabetes

Wnt7a promotes wound healing by regulation of angiogenesis and inflammation: Issues on and obesity.

Diabetic skin heals wounds poorly. Though obesity is the common risk factor of , few studies have investigated its effects on wound healing.This study aimed to evaluate the morphology and possible mechanism of human umbilical vein endothelial cells (HUVEC-C) in response to different levels of glucose and , and explore the role of Wnt7a in wound healing.The functional changes of HUVEC-C and mRNA expression of Wnt signaling were determined by analyzing cell viability, migration, tube formation and rt-PCR in gradients of glucose and . Recombinant Wnt7a protein was injected around wounds made on streptozotocin (STZ) -induced diabetic rats with (HF) or without (DM) high-fat diet. Angiogenesis and inflammatory statement were mainly analyzed by immunohistochemistry, ELISA, cytometry and Western blotting.The expression of Wnt7a significantly decreased in high Glc/PA cultured cells or DM and HF wounded rats. Impaired wound healing was also observed in DM and HF groups. The healing rate significantly accelerated after localized injection with Wnt7a at d10. Moreover, the expression of CD31, eNOS phosphorylation and NO were increased; the reduction of local neutrophils influx, ICAM-1 and IL-6/8 expression levels were obvious especially in diabetic with obesity rats at d10 after Wnt7a treatment.This study indicates the potential role of Wnt7a, which is beneficial for regeneration of damaged vessels, moderation of inflammatory statement in diabetic wound healing with or without obesity, thus demonstrating its possible utility as a topical administration to promote healing rate.Copyright © 2018. Published by Elsevier B.V.

Keyword: diabetes

Consumption of a high fat diet promotes protein O-GlcNAcylation in mouse retina via NR4A1-dependent GFAT2 expression.

The incidence of type 2 , the most common cause of diabetic retinopathy (DR), is rapidly on the rise in developed countries due to overconsumption of calorie rich diets. Using an animal model of diet-induced obesity/pre-, we evaluated the impact of a diet high in saturated fat (HFD) on O-GlcNAcylation of retinal proteins, as dysregulated O-GlcNAcylation contributes to diabetic complications and evidence supports a role in DR. Protein O-GlcNAcylation was increased in the retina of mice fed a HFD as compared to littermates receiving control chow. Similarly, O-GlcNAcylation was elevated in retinal Müller cells in culture exposed to the saturated fatty palmitate or the ceramide analog Cer6. One potential mechanism responsible for elevated O-GlcNAcylation is increased flux through the hexosamine biosynthetic pathway (HBP). Indeed, inhibition of the pathway\'s rate-limiting enzyme glutamine-fructose-6-phosphate amidotransferase (GFAT) prevented Cer6-induced O-GlcNAcylation. Importantly, expression of the mRNA encoding GFAT2, but not GFAT1 was elevated in both the retina of mice fed a HFD and in retinal cells in culture exposed to palmitate or Cer6. Notably, expression of nuclear receptor subfamily 4 group A member 1 (NR4A1) was increased in the retina of mice fed a HFD and NR4A1 expression was sufficient to promote GFAT2 mRNA expression and O-GlcNAcylation in retinal cells in culture. Whereas palmitate or Cer6 addition to culture medium enhanced NR4A1 and GFAT2 expression, chemical inhibition of NR4A1 transactivation repressed Cer6-induced GFAT2 mRNA expression. Overall, the results support a model wherein HFD increases retinal protein O-GlcNAcylation by promoting NR4A1-dependent GFAT2 expression.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Observational clinical and nerve conduction study on effects of a nutraceutical combination on painful diabetic distal symmetric sensory-motor neuropathy in patients with type 1 and type 2.

Painful distal symmetric polyneuropathy (pDSPN) is one of the most common and invalidating complications of , both of type 1 and type 2. Mechanisms responsible for the occurrence of the pDSPN are multifactorial and involve metabolic pathways regulating inflammation, microvessel circulation, axonal degeneration and so on. Several therapeutic approaches have been proposed to treat pain and each of them showed positive effects associated to drug-related side effects.Twenty-five consecutive patients with diagnosis of and pDSPN and tried to manage pain with a dietary supplement composed of a mixture of natural extracts (β-caryophyllene, myrrh, carnosic ) and PEA. This is a nutraceutical with potential multiple effects on metabolic, pain and vascular compartments, a profile considered useful in pDSPN. Patients were enrolled and polyneuropathy evaluated by means of nerve conduction study. Pain was assessed using VAS score scale and MNSI. Each patient was evaluated at T0 (time of enrollment) and at T1 (after 16 weeks of treatment).Supplement administration was well tolerated and induced unexpectedly significant amelioration of polyneuropathy with increase amplitude and reduction of pain. No side effects were reported.This fixed combination could well be considered as a potential nutraceutical option to manage pDSPN in diabetic patients.

Keyword: diabetes

PAQR3 regulates phosphorylation of FoxO1 in insulin-resistant HepG2 cells via NF-κB signaling pathway.

Insulin resistance is a significant feature of type 2 and glucose and lipid metabolism disorders. Activation of NF-κB signaling pathway plays an important role in the formation of insulin resistance. FoxO1 plays a major role in regulating glucose and lipid metabolism, as well as insulin signaling pathway. Previous studies have shown that Progestin and AdipoQ Receptor 3 (PAQR3) suppresses the activity of PI3K/Akt, which is an upstream pathway of FoxO1, and additionally promotes the pathological process of diabetic renal inflammatory fibrosis via activating NF-κB pathway. On this basis, it has caused us great concern whether NF-κB is involved in PAQR3 regulation of FoxO1 under insulin resistance. In this study, we aimed to investigate whether PAQR3 regulates phosphorylation of FoxO1 via NF-κB pathway in (PA)-induced insulin-resistant HepG2 cells, thereby causing glucose and lipid metabolism disorders. We found that PA stimulation and PAQR3 overexpression decreased the phosphorylation of FoxO1 and the expressions of glucokinase (GCK) and low density lipoprotein receptor (LDLR), in addition, promoted the nuclear accumulation of NF-κB. Inhibition of NF-κB pathway increased the phosphorylation of FoxO1 and the expressions of GCK and LDLR which were downregulated by PA stimulation and PAQR3 overexpression. Taken together, in PA-induced insulin-resistant HepG2 cells, PAQR3 might regulate the phosphorylation of FoxO1 and the expressions of GCK and LDLR through NF-κB pathway, thereby regulating the glucose and lipid metabolism disorders induced by insulin resistance.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic β-cells.

Free fatty receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic β-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated insulin secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of β-cell function. FFAR1 plays a key role in the development of type 2 (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of β-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic β-cells. Key players of the insulin signaling pathway, such as mTOR, Akt, IRS-1, and the insulin receptor (INSR1β), were selected as candidates to be analyzed under lipotoxic conditions.We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1β and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1β mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus.In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in β-cells under lipotoxic conditions.

Keyword: diabetes

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose metabolism in the obese mice.

Keyword: diabetes

Nutraceutical Potential of in Metabolic Syndrome.

L. is a well-known fruit worldwide, and its highest production occurs in tropical and subtropical regions. The pulp contains vitamins A, C, and E, B complex vitamins, such as pantothenic and folate, and minerals, such as magnesium and potassium, as well as food fibers. Phenolic compounds, such as benzyl isothiocyanate, glucosinolates, tocopherols (α and δ), β-cryptoxanthin, β-carotene and carotenoids, are found in the seeds. The oil extracted from the seed principally presents oleic fatty followed by , linoleic and stearic acids, whereas the leaves have high contents of food fibers and polyphenolic compounds, flavonoids, saponins, pro-anthocyanins, tocopherol, and benzyl isothiocyanate. Studies demonstrated that the nutrients present in its composition have beneficial effects on the cardiovascular system, protecting it against cardiovascular illnesses and preventing harm caused by free radicals. It has also been reported that it aids in the treatment of and in the reduction of cholesterol levels. Thus, both the pulp and the other parts of the plant (leaves and seeds) present antioxidant, anti-hypertensive, hypoglycemic, and hypolipidemic actions, which, in turn, can contribute to the prevention and treatment of obesity and associated metabolic disorders.

Keyword: diabetes

Erythrocyte Saturated Fatty Acids and Incident Type 2 in Chinese Men and Women: A Prospective Cohort Study.

The association between circulating saturated fatty acids (SFAs) and incident type 2 (T2D) is reported in Western populations with inconsistent results, while evidence from Asian populations is scarce. We aimed to examine the associations between erythrocyte SFAs and incident T2D in a Chinese population. Between 2008 and 2013, a total of 2683 participants, aged 40⁻75 years, free of were included in the present analyses. Incident T2D cases were ascertained during follow-up visits. Gas chromatography was used to measure erythrocyte fatty acids at baseline. The Cox proportional hazards model was used to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs). During 13,508 person years of follow-up, 216 T2D cases were identified. Compared with the first quartile, multivariable-adjusted HRs (95% CIs) of the fourth quartile were 1.20 (0.82⁻1.76; = 0.242) for myristic (14-carbon tail, zero double bonds; 14:0), 0.69 (0.48⁻0.99; = 0.080) for (16:0), 1.49 (1.02⁻2.19; = 0.047) for stearic (18:0), 1.46 (1.00⁻2.12; = 0.035) for arachidic (20:0), 1.48 (0.99⁻2.22; = 0.061) for behenic (22:0), and 1.08 (0.74⁻1.56; = 0.913) for lignoceric (24:0). Our findings indicate that individual erythrocyte SFAs are associated with T2D in different directions, with 18:0 and 20:0 SFAs positively associated with the risk, whereas no convincing inverse association for 16:0 SFAs.

Keyword: diabetes

Phlorizin Exerts Direct Protective Effects on (PA)-Induced Endothelial Dysfunction by Activating the PI3K/AKT/eNOS Signaling Pathway and Increasing the Levels of Nitric Oxide (NO).

BACKGROUND Sodium glucose transporter-2 inhibitors are the newest antidiabetic drugs that seem to be cardioprotective and can prevent type 2 in patients with high cardiovascular risks. Previous clinical trials have shown that these inhibitors can alleviate endothelial dysfunction, but the mechanism of action remains unknown. How SGLT inhibitor influences the release of NO in PA-induced HUVECs has never been reported. MATERIAL AND METHODS To explore the potential effects of the endothelial-protective mechanism of phlorizin and its impact on nitric oxide (NO), human umbilical vein endothelial cells (HUVECs) were incubated with (PA) and then treated with phlorizin. Western blotting was performed to assess the phosphorylation of AKT, eNOS, and IRS-1. To further explore potential targets, siRNA transfection was used to demonstrate the role of SGLT1 and SGLT2. RESULTS Phlorizin suppressed the expression of SGLT1 and SGLT2, activated the PI3K/AKT/eNOS signaling pathway, increased the output of NO, and promoted the consumption of glucose in PA-induced HUVECs. Through demonstrating siRNA suppression of the expression of SGLT1 and SGLT2 in PA-induced HUVECs, this study provides a new understanding of the mechanism behind SGLT1 and SGLT2. CONCLUSIONS Our data demonstrate that phlorizin ameliorates the endothelial dysfunction link with the activation of the PI3K/AKT/eNOS signaling pathway and augmentation of the release of NO, partially through suppressing the expression of SGLT1 and SGLT2 in PA-induced HUVECS.

Keyword: diabetes

Secreted Wnt6 mediates -associated centrosome amplification via its receptor FZD4.

We have recently published that type 2 promotes cell centrosome amplification via up-regulation of ROCK1 and 14-3-3σ. The study further investigated the molecular mechanisms underlying the -associated centrosome amplification. We found that treatment of cells with high glucose, insulin and increased the intracellular and the extracellular protein levels of Wnt6 as well as the cellular level of β-catenin. The treatment also activated β-catenin and promoted its nuclear translocation. Treatment of cells with siRNA species for Wnt6, FZD4 or β-catenin as well as introduction of antibodies against Wnt6 or FZD4 to the cell culture medium could all attenuate the treatment-triggered centrosome amplification. Moreover, we showed that secreted Wnt6-FZD4-β-catenin was the signaling pathway which was upstream of ROCK1 and 14-3-3σ. We also found that AGEs were also able to increase the cellular and extracellular levels of Wnt6, the cellular protein level of β-catenin, and centrosome amplification. Treatment of the cells with siRNA species for Wnt6 or FZD4 as well as introduction of antibodies against Wnt6 or FZD4 to the cell culture could all inhibit the AGEs-elicited centrosome amplification. In colon tissues from diabetic mice model, the protein levels of Wnt6 and 14-3-3σ were increased. In conclusion, our results showed that the pathophysiological factors in type 2 , including AGEs, were able to induce centrosome amplification. It is suggested that secreted Wnt6 binds to FZD4 to activate the canonical Wnt6 signaling pathway which is upstream of ROCK1 and 14-3-3σ, which is the cell signaling pathway underlying the -associated centrosome amplification.

Keyword: diabetes

Risk of associated with fatty acids in the de novo lipogenesis pathway is independent of insulin sensitivity and response: the Insulin Resistance Atherosclerosis Study (IRAS).

To examine the associations of fatty acids in the de novo lipogenesis (DNL) pathway, specifically myristic (14:0), (16:0), palmitoleic (c16:1\u2009n-7), myristoleic (c14:1n5), stearic (18:0) and oleic (c18:1\u2009n-9), with 5-year risk of type 2 . We hypothesized that DNL fatty acids are associated with risk of type 2 independent of insulin sensitivity.We evaluated 719 (mean age 55.1±8.5 years, 44.2% men, 42.3% Caucasians) participants from the Insulin Resistance Atherosclerosis Study. Multivariable logistic regression models with and without adjustment of insulin sensitivity were used to assess prospective associations of DNL fatty acids with incident type 2 .Type 2 incidence was 20.3% over 5\u2009years. In multivariable regression models, , palmitoleic, myristic, myristoleic and oleic acids were associated with increased risk of type 2 (p<0.05). had the strongest association (OR per standard unit of 1.46; 95%\u2009CI 1.23 to 1.76; p<0.001), which remained similar with addition of insulin sensitivity and acute insulin response (AIR) to the model (OR 1.36; 95%\u2009CI 1.09 to 1.70, p=0.01). Oleic and palmitoleic acids were also independently associated with incident type 2 . In multivariable models, ratios of fatty acids corresponding to stearoyl CoA desaturase-1 and Elovl6 enzymatic activity were significantly associated with risk of type 2 independent of insulin sensitivity and AIR.We observed associations of DNL fatty acids with type 2 incidence independent of insulin sensitivity.

Keyword: diabetes

N-3 Polyunsaturated Fatty Acids Decrease Long-Term Diabetic Risk of Offspring of Gestational Rats by Postponing Shortening of Hepatic Telomeres and Modulating Liver Metabolism.

The long-term influence of gestational (GDM) on offspring and the effect of omega-3 polyunsaturated fatty acids (n-3 PUFA) on GDM offspring are poorly understood. We studied the long-term diabetic risk in GDM offspring and evaluated the effect of n-3 PUFA intervention. Healthy offspring rats were fed standard diet (soybean oil) after weaning. GDM offspring were divided into three groups: GDM offspring (soybean oil), n-3 PUFA adequate offspring (fish oil), and n-3 PUFA deficient offspring (safflower oil), fed up to 11 months old. The diabetic risk of GDM offspring gradually increased from no change at weaning to obvious impaired glucose and insulin tolerance at 11 months old. N-3 PUFA decreased oxidative stress and inflammation in the liver of older GDM offspring. There was a differential effect of n-3 PUFA and n-6 PUFA on hepatic telomere length in GDM offspring. Non-targeted metabolomics showed that n-3 PUFA played a modulating role in the liver, in which numerous metabolites and metabolic pathways were altered when GDM offspring grew to old age. Many metabolites were related to risk, such as α-linolenic , , ceramide, oxaloacetic , tocotrienol, tetrahydro-11-deoxycortisol, andniacinamide. In summary, GDM offspring exhibited obvious risk at old age, whereas n-3 PUFA decreased this risk.

Keyword: diabetes

Dietary saturated fatty type impacts obesity-induced metabolic dysfunction and plasma lipidomic signatures in mice.

Saturated fatty (SFA) intake is associated with obesity, insulin resistance, and hepatic steatosis, but scant work examines the impact of SFA type upon these outcomes. We tested the hypothesis that an obesogenic diet prepared with medium chain SFA (MCSFA), mostly as lauric -derived from coconut oil, reduces obesity-induced outcomes compared to obesogenic diets prepared with increasing amounts long chain SFA (LCSFA), primarily . Mice were fed (16 weeks) a control, low fat diet or obesogenic diets prepared with differing content of MCSFA or LCSFA in which polyunsaturated and monounsaturated fatty acids (PUFA; MUFA) were kept constant. Inclusion of MCSFA in an obesogenic diet prevented hepatic lipid accumulation and lowered indices of insulin resistance. Obesogenic diets reduced hepatic levels of de novo lipogenesis proteins (SCD1 and FASN) but elevated the adipose levels of mRNA for the pro-inflammatory markers Mcp-1 and Tnfα. Lipidomic analysis of plasma indicated that MCSFA intake resulted in a different lipidomic signature than LCSFA intake, prevented elevation of pro-inflammatory ceramides, but elevated concentrations of some lipids associated with elevated cardiovascular disease risk. Intake of the obesogenic diets in an SFA-type dependent manner elevated plasma concentrations of several phosphatidylcholine (PC) lipids having the long chain PUFA (LCPUFA) arachidonic (ARA) and docosahexaenoic (DHA), altered phospholipid ethers, and changed the triacylglyceryl environments of these LCPUFA. Our data indicate that (1) MCSFA reduce the severity of some obesogenic co-morbidities, (2) SFA-type modulates lipidomic signatures associated with cardiovascular disease and , and (3) dietary SFA type impacts LCPUFA metabolism.Published by Elsevier Inc.

Keyword: diabetes

Hybrid lipids, peptides, and lymphocytes: new era in type 1 research.

Type 1 (T1D) results from autoimmune destruction of insulin-producing β cells in islets of Langerhans. Many genetic and immunological insights into autoimmune disease pathogenesis were initially uncovered in the context of T1D and facilitated by preclinical studies using the nonobese diabetic (NOD) mouse model. Recently, the study of T1D has led to the discovery of fatty esters of hydroxyl fatty acids (FAHFAs), which are naturally occurring hybrid peptides that modulate inflammation and pathogenesis, and a hybrid lymphocyte that expresses both B and T cell receptors. esters of hydroxy stearic acids (PAHSAs) are the most extensively studied FAHFA. In this issue of the JCI, Syed et al. have shown that PAHSAs both attenuate autoimmune responses and promote β cell survival in NOD mice. Given the lack of effective T1D therapies and the paucity of known side effects of PAHSAs, this lipid may have therapeutic potential for individuals at risk for or newly diagnosed with T1D.

Keyword: diabetes

PPARγ promotes -associated centrosome amplification via increasing the expression of SKA1 directly at the transcriptional level.

We have\xa0recently published that type 2 can induce cell centrosome amplification due to the action of high glucose, , and insulin, and ROCK1 and 14-3-3σ are signal mediators. In this study, we further investigated the molecular mechanisms of the centrosome amplification in colon cancer HCT116 cells. Treatment of the cells with high glucose, , and insulin increased the expression of peroxisome proliferator-activated receptor γ (PPARγ) as well as the spindle and kinetochore associated protein 1 (SKA1), knockdown of each of which resulted in the inhibition of the treatment-triggered centrosome amplification. Knockdown of PPARγ inhibited the treatment-evoked increase in the SKA1 level, whereas knockdown of SKA1 did not modify the treatment-increased PPARγ level. We found a predicted binding site for PPARγ in the promoter region of the SKA1 gene from the JASPAR database. Experimental results showed that the treatment increased the messenger RNA level of SKA1, which could be inhibited by PPARγ chemical inhibitor or small interfering RNA. Moreover, we were able to show that PPARγ could bind to the binding site in the SKA1 gene promoter, which was increased by the experimental treatment. In conclusion, it is suggested that the pathophysiological factors in type 2 , high glucose, , and insulin, induce the cell centrosome amplification through the PPARγ-SKA1 pathway, in which PPARγ increases the expression of SKA1 via directly enhancing the SKA1 gene transcription.© 2019 Wiley Periodicals, Inc.

Keyword: diabetes

Peroxidase expression is decreased by palmitate in cultured podocytes but increased in podocytes of advanced diabetic nephropathy.

High levels of serum free fatty acids (FFAs) are associated with lipotoxicity and type 2 . (PA) is the predominant circulating saturated FFA. PA induces mitochondrial superoxide and hydrogen peroxide (H O ) generation in cultured podocytes. To elucidate the role of PA in antioxidant defense systems in diabetic nephropathy (DN), cultured podocytes were exposed to 250\u2009μM PA for 1-24\u2009hr, and protein expressions of catalase, peroxiredoxins (Prxs), and glutathione peroxidase (GPx) were examined by western blot analysis. PA induced an early transient increase in the Prx1, Prx2, and GPx1 levels in podocytes, but not catalase. Long-term exposure of PA to podocytes significantly decreased the protein levels of Prx1, Prx2, GPx1, and catalase. Coincubation of PA-treated cells with oleic , however, restored the expression of these proteins. In advanced human diabetic glomeruli, H O generation was elevated as shown by increased fluorescence of dichlorofluorescein. Strong immunostaining for Prx1, Prx2, GPx1, and catalase was observed in the podocytes of advanced human DN, wherein transforming growth factor-β1 staining was also positive. These results suggest that podocytes are susceptible to PA-induced oxidative damage with impaired peroxidase activity and that peroxidases have futile antioxidant effects in the podocytes in the late stages of DN. Given this, PA-induced podocyte injury via inadequate peroxidase response to H O appears to play an important role in the pathogenesis of DN.© 2018 Wiley Periodicals, Inc.

Keyword: diabetes

Lipid profile changes in erythrocyte membranes of women with diagnosed GDM.

Gestational (GDM) is a glucose intolerance that begins or is first recognized during pregnancy. It is currently a growing health problem worldwide affecting from 1% to 14% of all pregnant women depending on racial and ethnic group as well as the diagnostic and screening criteria. Our preliminary study aimed at investigating the erythrocyte membrane fatty profiles of pregnant women, in particular with diagnosed with gestational (GDM), and with normal glucose tolerant (NGT) pregnant women as a control group. The study group comprised 43 pregnant women, 32 of whom were diagnosed with GDM according to the WHO criteria, and 11 with normal glucose tolerance. The erythrocyte membrane phospholipids were obtained according to the Folch extraction procedure. Fatty acids (FA) were analyzed by gas chromatography (GC) as the corresponding fatty methyl esters (FAME). A cluster of 14 fatty acids identified contained >98% of the recognized peaks in the GC analysis. The analysis of fatty acids from erythrocytes revealed important differences between GDM and NGT women in the third trimester, and the results were correlated with biochemical data. Among the 14 measured FA representing the membrane lipidomic profile, the levels of three saturated FA (myristic, , stearic acids) tended to decrease in GDM patients, with the percentage content of stearic significantly changed. The relative content of monounsaturated fatty acids (MUFA) tended to increase, in particular the oleic and vaccenic contents were significantly increased in erythrocyte membranes of the GDM group in comparison with the NGT group. The GDM group demonstrated higher sapienic levels (+29%) but this change was not statistically significant. This study revealed association between an impaired cis-vaccenic concentration in erythrocytes membrane and GDM development. No significant changes of polyunsaturated fatty acids (PUFA) were observed in GDM and NGT erythrocytes. We postulate, basing on the differences between the GDM and NGT lipidomic profiles, that stearic and cis-vaccenic acids can be considered as dual biomarkers of specific SFA-MUFA conversion pathway, involving the coupling of delta-9 desaturase and elongase enzymes. Our results indicate that the SFA-MUFA families may be involved in the pathophysiology of metabolic diseases such as GDM, but the further studies are needed to confirm our hypothesis. In conclusion, the erythrocyte membranes of GDM women undergo remodeling resulting in abnormal fatty profiles, which are reflection of the long-term status of organism and can have great impact on both the mother and her offspring.

Keyword: diabetes

Vernonia amygdalina Delile extract inhibits the hepatic gluconeogenesis through the activation of adenosine-5\'monophosph kinase.

It has been reported that Vernonia amygdalina Delile(VA) presents an anti-diabetic effect, and the effect of VA on lowering glucose is formulated via suppressing the expression of the key hepatic gluconeogenesis enzyme. Therefore, we further explored the probable mechanism of VA on dismissing hepatic gluconeogenesis through the activation of adenosine-5\' monophosphate kinase (AMPK) in vivo and in vitro.We developed type 2 diabetic mice with STZ and oral administration with VA (50\u202fmg/kg, 100\u202fmg/kg and 150\u202fmg/kg) once a day for 6 weeks. Fasting blood glucose (FBG), fasting insulin (FINS) and oral glucose tolerance tests (OGTT) were conducted. The expression levels of AMPK, phosphoenolpyruvate carboxykinase (PEPCK) and Glucose-6-phosphatase (G6Pase) proteins in live were evaluated by western blot. Then, we further explored the mechanism of VA on hepatic gluconeogenesis in vitro experiments. Glucose production and the expression of AMPK, PEPCK and G6Pase proteins were detected after VA treatment with the presence of the AMPK inhibitor Compound C.VA reduced FBG and caused a significant improvement in glucose tolerance and insulin resistance (HOMA-IR) in STZ-induced mice. VA inhibited the elevated expression of gluconeogenesis key enzymes (PEPCK and G6Pase) and up-regulated AMPK activity in liver. In (PA)-induced HepG2 cells, VA decreased glucose production and the expression of PEPCK and G6Pase proteins, also activated AMPK pathway. The effects of VA on gluconeogenesis could be reversed by Compound C.These results reveal that VA suppresses hepatic gluconeogenesis at least partially through activating the AMPK.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Effect of Hyperbaric Oxygen Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Patients: A Pilot Study.

Metabolic changes in insulin-dependent (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the lipid profile and altering FA composition.Copyright © 2019 Canadian Association. Published by Elsevier Inc. All rights reserved.

Keyword: diabetes

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high-fat diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and liver. PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free fatty acids during the clamp with Intralipid infusion reduced PAHSAs\' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial metabolic effects.

Keyword: diabetes

GCN2 deficiency ameliorates cardiac dysfunction in diabetic mice by reducing lipotoxicity and oxidative stress.

Excessive myocardial lipid accumulation is a major feature of diabetic cardiomyopathy (DCM). Although general control nonderepressible 2 (GCN2) has been identified as a sensor of amino availability, it also functions as an important regulator of hepatic lipid metabolism. Our previous studies have reported that GCN2 promotes pressure overload or doxorubicin-induced cardiac dysfunction by increasing cardiomyocyte apoptosis and myocardial oxidative stress. However, the impact of GCN2 on the development of DCM remains unclear. In this study, we investigated the effect of GCN2 on DCM in type 1 and type 2 animal models. After streptozotocin (STZ) or high-fat diet (HFD) plus low-dose STZ treatments, GCN2 mice developed less cardiac dysfunction, hyperlipidemia, myocardial hypertrophy, fibrosis, lipid accumulation, oxidative stress, inflammation and apoptosis compared with wild-type (WT) mice. In diabetic hearts, GCN2 deficiency attenuated the upregulation of peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ), the phosphorylation of eIF2α and the induction of activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), as well as the reduction of Bcl-2. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, high glucose or -induced cell death, oxidative and endoplasmic reticulum stress and lipid accumulation in H9C2 cells. Collectively, our data provide evidence that GCN2 deficiency protects cardiac function by reducing lipid accumulation, oxidative stress and cell death. Our findings suggest that strategies to inhibit GCN2 activity in the heart may be novel approaches for DCM therapy.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Synthesis and biological evaluations of marine oxohexadecenoic acids: PPARα/γ dual agonism and anti-diabetic target gene effects.

Obesity and associated disorders such as metabolic syndrome and type 2 (T2D) have reached epidemic proportions. Several natural products have been reported as Peroxisome Proliferator-Activated Receptor (PPAR) agonists, functioning as lead compounds towards developing new anti-diabetic drugs due to adverse side effects of existing PPAR drugs. We recently isolated and identified (7E)-9-oxohexadec-7-enoic (1) and (10E)-9-oxohexadec-10-enoic (2) from the marine algae Chaetoceros karianus. Herein we report the total synthesis, pharmacological characterization, and biological evaluations of these naturally occurring oxo-fatty acids (oFAs). The syntheses of 1 and 2 afforded sufficient material for extensive biological evaluations. Both oFAs show an appreciable dose-dependent activation of PPARα and -γ, with EC values in the micromolar range, and an ability to regulate important PPAR target genes in hepatocytes and adipocytes. Moreover, both 1 and 2 are able to drive adipogenesis when evaluated in the Simpson-Golabi-Behmel syndrome (SGBS) pre-adipocyte cell model, but with lowered expression of adipocyte markers and reduced lipid accumulation compared to the drug rosiglitazone. This seems to be caused by a transient upregulation of PPARγ and C/EBPα expression. Importantly, whole transcriptome analysis shows that both compounds induce anti-diabetic gene programs in adipocytes by upregulating insulin-sensitizing adipokines and repressing pro-inflammatory cytokines.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: diabetes

Alkannin Inhibited Hepatic Inflammation in Diabetic Db/Db Mice.

The current study was designed to investigate the protective role of alkannin (ALK) on liver injury in diabetic C57BL/KsJ-db/db mice and explore its potential mechanisms.An oral glucose tolerance test (OGTT) was performed. The levels of insulin, alanine aminotransferase (ALT), aspartate aminotransaminase (AST), total cholesterol (TC) and triglyceride (TG) were determined by commercial kits. The pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α were determined by ELISA. The levels of the ROCK/NF-κB pathway were determined by Western blotting.The contents of pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α were inhibited by ALK, metformin or fasudil in diabetic db/db mice. Further, Western blotting analysis showed that the expression of Rho, ROCK1, ROCK2, p-NF-κBp65, and p-IκBα was significantly reversed by ALK treatment. In human hepatic HepG2 cells, the hepatoprotective effects of ALK were further characterized. With response to -challenge, increased amounts of insulin, ALT, AST, TG, and TC were observed, whereas ALK pretreatment significantly inhibited their leakage in HepG2 cells without appreciable cytotoxic effects. The inflammation condition was recovered with ALK treatment as shown by changes of IL-1β, IL-6 and TNF-α. Further, Western blotting analysis also suggested that ALK improves hepatic inflammation in a Rho-kinase pathway.The present study successfully investigated the role of Rho-kinase signalling in diabetic liver injury. ALK exhibited hepatoprotective effects in diabetic db/db mice, and it might act through improving hepatic inflammation through the Rho-kinase pathway.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Astragalosides IV protected the renal tubular epithelial cells from free fatty acids-induced injury by reducing oxidative stress and apoptosis.

Renal tubular injury is associated with the development of diabetic nephropathy (DN) and the end-stage renal disease (ESRD). Free fatty acids (FFAs)-associated lipotoxicity contributes to injury of proximal renal tubular epithelial (HK-2) cells in . (PA) which is the most abundant saturated fatty in FFAs is closely associated with the gradual decline of renal function. Astragalosides IV (AS-IV) has a variety of pharmacological effects such as anti-inflammation and anti-oxidation. In the current study, we investigated the effects of AS-IV on PA-induced apoptosis of HK-2 cells and the underlying mechanisms. The results showed that AS-IV (10, 20, 40\u2009μmol/L) could alleviate PA-induced apoptosis of HK-2 cells. We found that AS-IV reduced the expression of Bax and cleaved-caspase3, but increased the expression of Bcl-2 and phosphorylated Nrf2 in HK-2 cells. Moreover, AS-IV reduced the level of reactive oxygen species (ROS) in the cells. Our study suggests that AS-IV could protect against PA-induced apoptosis in HK-2 cells by inhibiting ROS generation and apoptotic protein expression. This study may provide a new theoretical option for the patients with type 2 .Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: diabetes

Nucleophosmin/B23 contributes to hepatic insulin resistance through the modulation of NF-κB pathway.

Nucleophosmin (NPM)/B23 is an important nucleolar phosphoprotein involved in the regulation of assorted cellular signaling pathways. In the present study, we revealed a critical role of NPM in liver insulin resistance. NPM is markedly upregulated in insulin-resistant liver tissues and (PA)-exposed HepG2 cells both at mRNA and protein levels. Ectopic expression of NPM in hepatocytes aggravated PA-induced insulin resistance, lipid droplet accumulation, glucose intake impairment as well as the expression of gluconeogenic genes. Coinciding with these results, interference of NPM using small interfering RNA (siRNA) oligos ameliorated PA-induced insulin resistance, as revealed by increased phosphorylation of AKT and GSK3β following insulin treatment. As predicted, PA-triggered alterations in glucose intake and the expression of gluconeogenic enzymes were attenuated following NPM depletion. Finally, we showed that NPM plays an indispensible role in PA-induced activation of NF-κB pathway. Both of NF-κB p65 phosphorylation and nuclear translocation were impeded by NPM interference in PA-treated HepG2 cells. Taken together, these findings explicitly demonstrate that NPM participates in the development of liver insulin resistance, suggesting that NPM may serve as a potential therapeutic target of type 2 .Copyright © 2019. Published by Elsevier Inc.

Keyword: diabetes

Attenuation of Free Fatty -Induced Muscle Insulin Resistance by Rosemary Extract.

Elevated blood free fatty acids (FFAs), as seen in obesity, impair muscle insulin action leading to insulin resistance and Type 2 . Serine phosphorylation of the insulin receptor substrate (IRS) is linked to insulin resistance and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the energy sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract insulin resistance. We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle insulin resistance has never been examined. In the current study, we investigated the effect of RE in palmitate-induced insulin resistant L6 myotubes. Exposure of myotubes to palmitate reduced the insulin-stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the insulin-stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the insulin-stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell insulin resistance and further studies are required to explore its antidiabetic properties.

Keyword: diabetes

PAHSAs attenuate immune responses and promote β cell survival in autoimmune diabetic mice.

esters of hydroxy stearic acids (PAHSAs) are endogenous antidiabetic and antiinflammatory lipids. Here, we show that PAHSAs protect against type 1 (T1D) and promote β cell survival and function. Daily oral PAHSA administration to nonobese diabetic (NOD) mice delayed the onset of T1D and markedly reduced the incidence of T1D, whether PAHSAs were started before or after insulitis was established. PAHSAs reduced T and B cell infiltration and CD4+ and CD8+ T cell activation, while increasing Treg activation in pancreata of NOD mice. PAHSAs promoted β cell proliferation in both NOD mice and MIN6 cells and increased the number of β cells in NOD mice. PAHSAs attenuated cytokine-induced apoptotic and necrotic β cell death and increased β cell viability. The mechanism appears to involve a reduction of ER stress and MAPK signaling, since PAHSAs lowered ER stress in NOD mice, suppressed thapsigargin-induced PARP cleavage in human islets, and attenuated ERK1/2 and JNK1/2 activation in MIN6 cells. This appeared to be mediated in part by glucagon-like peptide 1 receptor (GLP-1R) and not the G protein-coupled receptor GPR40. PAHSAs also prevented impairment of glucose-stimulated insulin secretion and improved glucose tolerance in NOD mice. Thus, PAHSAs delayed the onset of T1D and reduced its incidence by attenuating immune responses and exerting direct protective effects on β cell survival and function.

Keyword: diabetes

PCNA and JNK1-Stat3 pathways respectively promotes and inhibits -associated centrosome amplification by targeting at the ROCK1/14-3-3σ complex in human colon cancer HCT116 cells.

We have recently reported that type 2 promotes centrosome amplification via enhancing the expression, biding, and centrosome translocation of rho-associated coiled-coil containing protein kinase 1 (ROCK1)/14-3-3σ complex in HCT116 cells. In the functional proteomic study, we further investigated the molecular pathways underlying the centrosome amplification using HCT116 cells. We found that treatment of HCT116 cells with high glucose, insulin, and triggered the centrosome amplification and increased the expressions of proliferating cell nuclear antigen (PCNA), nucleophosmin (NPM), and 14-3-3σ. Individual knockdown of PCNA, NPM, or 14-3-3σ inhibited the centrosome amplification. Knockdown of PCNA inhibited the treatment-increased expression of ROCK1, whereas knockdown of ROCK1 did not affect the PCNA expression. High glucose, insulin, and also increased the expressions of c-Jun N-terminal kinase-1 (JNK1) and signal transducer and activator of transcription 3 (Stat3), individual knockdown of which upregulated the treatment-increased expression of 14-3-3σ and promoted the centrosome amplification. In contrast, overexpression of JNK1 inhibited the centrosome amplification. Knockdown of Stat3 enhanced the centrosome translocation of 14-3-3σ. Moreover, we showed that knockdown of JNK1 inhibited the treatment-increased expression of Stat3. Knockdown of PCNA, JNK1, or Stat3 did not have an effect on NPM and vice versa. In conclusion, our results suggest that PCNA and JNK1-Stat3 pathways respectively promotes and feedback inhibits the centrosome amplification by targeting at the ROCK1/14-3-3σ complex, and NPM serves as an independent signal for the centrosome amplification.© 2018 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

Keyword: diabetes

Evidence for an alternative fatty desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated lipid metabolism that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty metabolism and, in particular, fatty desaturation. This suggests that many cancer cells contain an unexplored plasticity in their fatty metabolism. Here we show that some cancer cells can exploit an alternative fatty desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in fatty desaturation and constitutes an unexplored metabolic rewiring in cancers.

Keyword: diabetes

Cellular toxicity of dietary trans fatty acids and its correlation with ceramide and diglyceride accumulation.

High fatty (FA) levels are deleterious to pancreatic β-cells, largely due to the accumulation of biosynthetic lipid intermediates, such as ceramides and diglycerides, which induce ER stress and apoptosis. Toxicity of palmitate (16:0) and oleate (18:1 cis-Δ) has been widely investigated, while very little data is available on the cell damages caused by elaidate (18:1 trans-Δ) and vaccenate (18:1 trans-Δ), although the potential health effects of these dietary trans fatty acids (TFAs) received great publicity. We compared the effects of these four FAs on cell viability, apoptosis, ER stress, JNK phosphorylation and autophagy as well as on ceramide and diglyceride contents in RINm5F insulinoma cells. Similarly to oleate and unlike palmitate, TFAs reduced cell viability only at higher concentration, and they had mild effects on ER stress, apoptosis and autophagy. Palmitate increased ceramide and diglyceride levels far more than any of the unsaturated fatty acids; however, incorporation of TFAs in ceramides and diglycerides was strikingly more pronounced than that of oleate. This indicates a correlation between the accumulation of lipid intermediates and the severity of cell damage. Our findings reveal important metabolic characteristics of TFAs that might underlie a long term toxicity and hence deserve further investigation.Copyright © 2018. Published by Elsevier Ltd.

Keyword: diabetes

Complement Component C3 Is Highly Expressed in Human Pancreatic Islets and Prevents β Cell Death via ATG16L1 Interaction and Autophagy Regulation.

We show here that human pancreatic islets highly express C3, which is both secreted and present in the cytosol. Within isolated human islets, C3 expression correlates with type 2 (T2D) donor status, HbA1c, and inflammation. Islet C3 expression is also upregulated in several rodent models. C3 interacts with ATG16L1, which is essential for autophagy. Autophagy relieves cellular stresses faced by β cells during T2D and maintains cellular homeostasis. C3 knockout in clonal β cells impaired autophagy and led to increased apoptosis after exposure of cells to and IAPP. In the absence of C3, autophagosomes do not undergo fusion with lysosomes. Thus, C3 may be upregulated in islets during T2D as a cytoprotective factor against β cell dysfunction caused by impaired autophagy. Therefore, we revealed a previously undescribed intracellular function for C3, connecting the complement system directly to autophagy, with a broad\xa0potential importance in other diseases and cell types.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Liraglutide protects non-alcoholic fatty liver disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high-fat diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 (T2DM), has also been reported to decrease fatty degeneration of the liver. The purpose of this study is to explore whether liraglutide improves high-fat diet-induced non-alcoholic fatty liver disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the liver. After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg body weight) for four weeks, the liver, liver/body weight, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high-fat diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the liver of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high-fat diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Ceramide stearic to ratio predicts incident .

Ceramide lipids have a role in the development of insulin resistance, and risk of cardiovascular disease. Here we investigated four ceramides and their ratios to find the best predictors of incident .A validated mass-spectrometric method was applied to measure Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) from serum or plasma samples. These ceramides were analysed in a population-based risk factor study (FINRISK 2002, n\u2009=\u20098045), in a cohort of participants undergoing elective coronary angiography for suspected stable angina pectoris (Western Norway Coronary Angiography Cohort [WECAC], n\u2009=\u20093344) and in an intervention trial investigating improved methods of lifestyle modification for individuals at high risk of the metabolic syndrome (Prevent Metabolic Syndrome [PrevMetSyn], n\u2009=\u2009371). risk score models were developed to estimate the 10\xa0year risk of incident .Analysis in FINRISK 2002 showed that the Cer(d18:1/18:0)/Cer(d18:1/16:0) ceramide ratio was predictive of incident (HR per SD 2.23, 95% CI 2.05, 2.42), and remained significant after adjustment for several risk factors, including BMI, fasting glucose and HbA (HR 1.34, 95% CI 1.14, 1.57). The finding was validated in the WECAC study (unadjusted HR 1.81, 95% CI 1.53, 2.14; adjusted HR 1.39, 95% CI 1.16, 1.66). In the intervention trial, the ceramide ratio and risk scores significantly decreased in individuals who had 5% or more weight loss.The Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio is an independent predictive biomarker for incident , and may be modulated by lifestyle intervention.

Keyword: diabetes

Acute and Repeated Treatment with 5-PAHSA or 9-PAHSA Isomers Does Not Improve Glucose Control in Mice.

Fatty esters of hydroxylated fatty acids (FAHFAs) were discovered as a novel class of endogenous mammalian lipids whose profound effects on metabolism have been shown. In the current study, in\xa0vitro and in\xa0vivo the metabolic effects of two of these FAHFAs, namely -5- (or -9) -hydroxy-stearic (5- or 9-PAHSA, respectively) were profiled. In DIO mice fed with differentially composed low- or high-fat diets, acute and subchronic treatment with 5-PAHSA and 9-PAHSA alone, or in combination, did not significantly improve the deranged metabolic status. Neither racemic 5- or 9-PAHSA, nor the enantiomers were able to: (1) increase basal or insulin-stimulated glucose uptake in\xa0vitro, (2) stimulate GLP-1 release from GLUTag cells, or (3) induce GSIS in rat, mouse, or human islets or in a human pancreatic β cell line. Therefore, our data do not support the\xa0further development of PAHSAs or their derivatives for the control of insulin resistance and hyperglycemia.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Kaempferol alleviates -induced lipid stores, endoplasmic reticulum stress and pancreatic β-cell dysfunction through AMPK/mTOR-mediated lipophagy.

Kaempferol, a natural flavonoid, has the beneficial effects of preserving pancreatic β-cell mass and function, but its action on β-cell lipid metabolism still remains elusive. Recently, autophagy has been reported to play a major role in lipid metabolism in various cell types, but its role in pancreatic β-cell\'s lipid metabolism is rarely reported. Here, we investigated the role of kaempferol-induced autophagy in inhibition of lipid stores, ER stress and β-cell dysfunction in -challenged RIN-5F cells and isolated pancreatic islets. The lipid-lowering effect of kaempferol was determined by Oil Red O staining, triglyceride assay, BODIPY labeling, RT-PCR and immunoblot analysis of PLIN2 (the lipid droplet coat protein) expression. Further, the involvement of AMPK/mTOR-mediated lipophagy was established by pharmacological and genetic inhibitors of autophagy and AMPK. The co-localization studies of lipid droplets with autophagosomes/lysosomes by BODIPY-MDC-LysoTracker co-staining, LC3/BODIPY labeling and LC3/PLIN2 double immunolabeling further strengthened the findings. Kaempferol treatment exhibited decreased lipid stores and increased co-localization of lipid droplets with autophagosomes and lysosomes in -challenged β-cells. Moreover, inhibition of autophagy led to decreased co-localization and increased lipid droplets accumulation. Kaempferol-induced alleviation of ER stress and β-cell dysfunctions was established by immunoblot analysis of CHOP-10 (a key mediator of cell death in response to ER stress) and insulin content/secretion analysis respectively. Together, these findings suggest that kaempferol prevents ectopic lipid accumulation and ER stress, thus restoring β-cell function through AMPK-mediated lipophagy. The current data implies that kaempferol may be a potential therapeutic candidate to prevent obesity-linked diabetic complications.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Faster Protocol for Endogenous Fatty Esters of Hydroxy Fatty (FAHFA) Measurements.

Fatty esters of hydroxy fatty acids (FAHFAs) are a recently discovered class of endogenous lipids with antidiabetic and anti-inflammatory activities. Interest in these lipids is due to their unique biological activites and the observation that insulin-resistant people have lower esters of hydroxystearic (PAHSA) levels, suggesting that a FAHFA deficiency may contribute to metabolic disease. Rigorous testing of this hypothesis will require the measurement of many clinical samples; however, current analytical workflows are too slow to enable samples to be analyzed quickly. Here we describe the development of a significantly faster workflow to measure FAHFAs that optimizes the fractionation and chromatography of these lipids. We can measure FAHFAs in 30 min with this new protocol versus 90 min using the older protocol with comparable performance in regioisomer detection and quantitation. We also discovered through this optimization that oleic esters of hydroxystearic acids (OAHSAs), another family of FAHFAs, have a much lower background signal than PAHSAs, which makes them easier to measure. Our faster workflow was able to quantify changes in PAHSAs and OAHSAs in mouse tissues and human plasma, highlighting the potential of this protocol for basic and clinical applications.

Keyword: diabetes

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of , obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and liver function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 liver conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe liver inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: diabetes

A novel recombinant peptide INSR-IgG4Fc (Yiminsu) restores insulin sensitivity in experimental insulin resistance models.

Type 2 (T2DM) is a chronic degenerative endocrine and metabolic disease with high mortality and morbidity, yet lacks effective therapeutics. We recently generated a novel fusion peptide INSR-IgG4Fc, Yiminsu (YMS), to facilitate the high-affinity binding and transportation of insulin. Thus, the aim of the present study was to determine whether the novel recombinant peptide, YMS, could contribute to restoring insulin sensitivity and glycaemic control in insulin resistance models and revealing its underlying mechanism. (PA)-treated LO2 cells and high fat diet (HFD)-fed mice were treated with YMS. Therapeutic effects of YMS were measured using Western blotting, ELISA, qPCR, Histology and transmission electron microscopy. We observed that YMS treatment effectively improved insulin signaling in PA-treated LO2 cells and HFD-fed mice. Notably, YMS could significantly reduce serum levels of glucose, triglycerides, fatty acids and cholesterol without affecting the serum insulin levels. Moreover, our data demonstrated that YMS could restore glucose and lipid homeostasis via facilitating insulin transportation and reactivating PI3K/Akt signaling in both PA-treated cells and liver, gastrocnemius and brown fat of HFD-fed mice. Additionally, we noticed that the therapeutic effects of YMS was similar as rosiglitazone, a well-recognized insulin sensitizer. Our findings suggested that YMS is a potentially candidate for pharmacotherapy for metabolic disorders associated with insulin resistance, particularly in T2DM.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: diabetes

Myo-inositol alters 13C-labeled fatty metabolism in human placental explants.

We postulate that myo-inositol, a proposed intervention for gestational-, affects transplacental lipid supply to the fetus. We investigated the effect of myo-inositol on fatty- processing in human placental-explants from uncomplicated pregnancies. Explants were incubated with 13C-labeled , 13C-oleic- and 13C-docosahexaenoic- across a range of myo-inositol concentrations for 24 h and 48 h. The incorporation of labeled-fatty-acids into individual lipids was quantified by liquid-chromatography-mass-spectrometry. At 24 h, myo-inositol increased the amount of 13C- and 13C-oleic- labeled lipids (median fold-change relative to control=1). Significant effects were seen with 30 µM myo-inositol (physiological) for 13C--lysophosphatidylcholines (1.26) and 13C--phosphatidylethanolamines (1.17). At 48 h, myo-inositol addition increased 13C-oleic--lipids but decreased 13C- and 13C-docosahexaenoic- lipids. Significant effects were seen with 30 µM myo-inositol for 13C-oleic--phosphatidylcholines (1.25), 13C-oleic--phosphatidylethanolamines (1.37) and 13C-oleic--triacylglycerols (1.32) and with 100 µM myo-inositol for 13C-docosahexaenoic--triacylglycerols (0.78). Lipids labeled with the same 13C-fatty- showed similar responses when tested at the same time-point, suggesting myo-inositol alters upstream processes such as fatty- uptake or activation. Myo-inositol supplementation may alter placental lipid physiology with unknown clinical consequences.

Keyword: diabetes

Interesterified palm olein lowers postprandial glucose-dependent insulinotropic polypeptide response in type 2 .

We aim to investigate the postprandial effects of palm olein (PO) and chemically interesterified palm olein (IPO) with different proportions of at the sn-2 position using high oleic sunflower oil (HOS) as control fat on concentrations of gut hormones, glucose homeostasis, satiety, lipid and inflammatory parameters in type 2 diabetic (T2D) subjects.Using a randomised double-blind crossover design, 21 (men\u2009=\u20096, women\u2009=\u200915) T2D subjects consumed test meals (3.65\xa0MJ) consisting of a high fat muffin (containing 50\xa0g test fats provided as PO, IPO or HOS) and a milkshake. Postprandial changes in gut hormones, glucose homeostasis, satiety, lipid and inflammatory parameters after meals were analysed. Some of the solid fractions of the IPO were removed and thus the fatty composition of the PO and IPO was not entirely equal (PO vs IPO: palmitate 39.8 vs 38.7; oleate 43.6 vs 45.1). PO, IPO and HOS contained 9.7, 38.9 and 0.2\xa0g/100\xa0g total fatty acids of at the sn-2 position, respectively. At 37\xa0°C, IPO contained 4.2% SFC whereas PO and HOS were completely melted.Our novel observation shows that the incremental area under curve (iAUC) 0-6\xa0h of plasma GIP concentration was on average 16% lower following IPO meal compared with PO and HOS (P\u2009<\u20090.05) meals. Serum C-peptide concentrations exhibited a significant meal\u2009×\u2009gender interaction (P\u2009=\u20090.009). No differences between test meals were noted for other measurements.This study shows no adverse effect of interesterification on hormones associated with glucose homeostasis notably GLP-1 in T2D subjects.ClinicalTrials.gov . https://clinicaltrials.gov/ct2/show/.

Keyword: diabetes

Circulating Very-Long-Chain SFA Concentrations Are Inversely Associated with Incident Type 2 in US Men and Women.

Very-long-chain SFAs (VLCSFAs), such as arachidic (20:0), behenic (22:0), and lignoceric (24:0), have demonstrated inverse associations with cardiometabolic conditions, although more evidence is needed to characterize their relation with risk of type 2 (T2D). In addition, little is known regarding their potential dietary and lifestyle predictors.We aimed to examine the association of plasma and erythrocyte concentrations of VLCSFAs with incident T2D risk.We used existing measurements of fatty concentrations in plasma and erythrocytes among 2854 and 2831 participants in the Nurses\' Health Study (NHS) and Health Professionals Follow-Up Study (HPFS), respectively. VLCSFAs were measured using GLC, and individual fatty concentrations were expressed as a percentage of total fatty acids. Incident T2D cases were identified by self-reports and confirmed by a validated supplementary questionnaire. Cox proportional hazards regression was used to evaluate the association between VLCSFAs and T2D, adjusting for demographic, lifestyle, and dietary variables.During 39,941 person-years of follow-up, we documented 243 cases of T2D. Intakes of peanuts, peanut butter, vegetable fat, dairy fat, and /stearic (16:0-18:0) fatty acids were significantly, albeit weakly, correlated with plasma and erythrocyte VLCSFA concentrations (|rs|\xa0≤\xa00.19). Comparing the highest with the lowest quartiles of plasma concentrations, pooled HRs (95% CIs) were 0.51 (0.35, 0.75) for arachidic , 0.43 (0.28, 0.64) for behenic , 0.40 (0.27, 0.61) for lignoceric , and 0.41 (0.27, 0.61) for the sum of VLCSFAs, after multivariate adjustments for demographic, lifestyle, and dietary factors. For erythrocyte VLCSFAs, only arachidic and behenic concentrations were inversely associated with T2D risk.Our findings suggest that, in US men and women, higher plasma concentrations of VLCSFAs are associated with lower risk of T2D. More research is needed to understand the mechanistic pathways underlying these associations.Copyright © The Author(s) 2019.

Keyword: diabetes

The monounsaturated fatty oleate is the major physiological toxic free fatty for human beta cells.

Free fatty acids (FFAs) can cause glucose intolerance and . Lipotoxicity to the pancreatic beta cells is considered to be a major underlying cause for this phenomenon. The aim of this study was to analyse the toxicity profile of FFAs in the human EndoC-βH1 beta-cell line and to compare the results with isolated rat and human islets with special reference to the physiologically most prevalent FFAs (PA) and oleic (OA). Toxicity after a 2-day incubation with the different FFAs was analysed by the caspase-3 assay and confirmed by the propidium iodide and annexin V staining tests. The long-chain saturated PA (C16:0) and the monounsaturated OA (C18:1) were both toxic to human EndoC-βH1 beta cells and pseudoislets, as well as to rat islets, and, as confirmed in a pilot experiment, also to human islets. Furthermore, OA provided no protection against the toxicity of PA. Likewise, elaidic (EA, the trans isomer of OA; trans-OA) was significantly toxic, in contrast to the non-metabolisable analogues methylated PA (MePA) and methylated OA (MeOA). Fatty acids with a chain length\u2009\u2009< C16 were not toxic in EndoC-βH1 beta cells. Caspase-3 was also activated by linoleic (LA)(C18:2) but not by γ-linolenic (γ-LNA)(C18:3). Overall, only long-chain FFAs with chain lengths\u2009 >\u2009C14, which generate hydrogen peroxide in the peroxisomal beta-oxidation, were toxic. This conclusion is also supported by the toxicity of the branched-chain FFA pristanic , which is exclusively metabolised in the peroxisomal beta-oxidation. The lack of a protective effect of the monounsaturated fatty OA has important consequences for a beta-cell protective lipid composition of a diet. A cardioprotective diet with a high OA content does not fulfil this requirement.

Keyword: diabetes

Exogenous Hydrogen Sulfide Alleviates-Induced Intracellular Inflammation in HepG2 Cells.

Fatty acids induced hepatic inflammation plays an important role in nonalcoholic fatty liver disease (NAFLD) pathogenesis. Hydrogen sulfide (HS), an endogenous gasotransmitter, has been established to possess potent anti-inflammation in various human organs. However, the anti-inflammation property of HS in the fatty liver is still needed to further elucidate. Hence, this study aimed to investigate whether exogenous HS can protect hepatocytes against inflammation induced by (PA). HepG2 hepatocytes were exposed to PA for 24\u2009h to induce free fatty acids-induced inflammation. The cells were pretreated with NaHS (a donor of H2S) before exposure to PA. Cell viability, inflammatory cytokines (TNF-α, IL-6 and IL-1β), NLRP3 inflammasome and NF-κB were measured by a combination of MTT assay, ELISA, Western blot and Immunofluorescence. Here, we found that exogenous HS dose-dependently inhibited the expression of pro-inflammatory cytokines, NLRP3 inflammasome and activation of NF-κB signaling in PA-induced HepG2 cells. Thus, HS might be a candidate therapeutic agent against NAFLD.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: diabetes

20-HETE promotes glucose-stimulated insulin secretion in an autocrine manner through FFAR1.

The long-chain fatty receptor FFAR1 is highly expressed in pancreatic β-cells. Synthetic FFAR1 agonists can be used as antidiabetic drugs to promote glucose-stimulated insulin secretion (GSIS). However, the physiological role of FFAR1 in β-cells remains poorly understood. Here we show that 20-HETE activates FFAR1 and promotes GSIS via FFAR1 with higher potency and efficacy than dietary fatty acids such as , linoleic, and α-linolenic . Murine and human β-cells produce 20-HETE, and the ω-hydroxylase-mediated formation and release of 20-HETE is strongly stimulated by glucose. Pharmacological inhibition of 20-HETE formation and blockade of FFAR1 in islets inhibits GSIS. In islets from type-2 diabetic humans and mice, glucose-stimulated 20-HETE formation and 20-HETE-dependent stimulation of GSIS are strongly reduced. We show that 20-HETE is an FFAR1 agonist, which functions as an autocrine positive feed-forward regulator of GSIS, and that a reduced glucose-induced 20-HETE formation contributes to inefficient GSIS in type-2 .

Keyword: diabetes

Melatonin improves insulin resistance and hepatic steatosis through attenuation of alpha-2-HS-glycoprotein.

Melatonin plays an important role in regulating circadian rhythms. It also acts as a potent antioxidant and regulates glucose and lipid metabolism, although the exact action mechanism is not clear. The α2-HS-glycoprotein gene (AHSG) and its protein, fetuin-A (FETUA), are one of the hepatokines and are known to be associated with insulin resistance and type 2 . The aim of this study was to determine whether melatonin improves hepatic insulin resistance and hepatic steatosis in a FETUA-dependent manner. In HepG2 cells treated with 300\xa0μmol/L of , phosphorylated AKT expression decreased, and FETUA expression increased, but this effect was inhibited by treatment with 10\xa0μmol/L of melatonin. However, melatonin did not improve insulin resistance in FETUA-overexpressing cells, indicating that improvement in insulin resistance by melatonin was dependent on downregulation of FETUA. Moreover, melatonin decreased -induced ER stress markers, CHOP, Bip, ATF-6, XBP-1, ATF-4, and PERK. In addition, in the high-fat diet (HFD) mice, oral treatment with 100\xa0mg/kg/day melatonin for 10\xa0weeks reduced body weight gain to one-third of that of the HFD group and hepatic steatosis. Insulin sensitivity and glucose intolerance improved with the upregulation of muscle p-AKT protein expression. FETUA expression and ER stress markers in the liver and serum of HFD mice were decreased by melatonin treatment. In conclusion, melatonin can improve hepatic insulin resistance and hepatic steatosis through reduction in ER stress and the resultant AHSG expression.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: diabetes

The BACE1 product sAPPβ induces ER stress and inflammation and impairs insulin signaling.

β-secretase/β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a key enzyme involved in Alzheimer\'s disease that has recently been implicated in insulin-independent glucose uptake in myotubes. However, it is presently unknown whether BACE1 and the product of its activity, soluble APPβ (sAPPβ), contribute to lipid-induced inflammation and insulin resistance in skeletal muscle cells.Studies were conducted in mouse C2C12 myotubes, skeletal muscle from Bace1mice and mice treated with sAPPβ and adipose tissue and plasma from obese and type 2 diabetic patients.We show that BACE1 inhibition or knockdown attenuates palmitate-induced endoplasmic reticulum (ER) stress, inflammation, and insulin resistance and prevents the reduction in Peroxisome Proliferator-Activated Receptor γ Co-activator 1α (PGC-1α) and fatty oxidation caused by palmitate in myotubes. The effects of palmitate on ER stress, inflammation, insulin resistance, PGC-1α down-regulation, and fatty oxidation were mimicked by soluble APPβ in vitro. BACE1 expression was increased in subcutaneous adipose tissue of obese and type 2 diabetic patients and this was accompanied by a decrease in PGC-1α mRNA levels and by an increase in sAPPβ plasma levels of obese type 2 diabetic patients compared to obese non-diabetic subjects. Acute sAPPβ administration to mice reduced PGC-1α levels and increased inflammation in skeletal muscle and decreased insulin sensitivity.Collectively, these findings indicate that the BACE1 product sAPPβ is a key determinant in ER stress, inflammation and insulin resistance in skeletal muscle and gluconeogenesis in liver.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Chronic -induced lipotoxicity correlates with defective trafficking of ATP sensitive potassium channels in pancreatic β cells.

Lipotoxicity is associated with a high level of fatty accumulation in pancreatic β-cells. An overload of free fatty acids contributes to pancreatic β-cell apoptosis and dysfunction. Insulin secretion involves sequential ionic events upon glucose stimulation. ATP sensitive potassium (K) channels serve as glucose sensors and effectively initiate glucose-stimulated insulin secretion. This study investigated the effects of lipotoxicity on the trafficking of K channels in pancreatic β cells using chronic -injected mice and treated insulinoma cells. The chronic -injected mice displayed type II diabetic characteristics. The pancreatic sections of these mice exhibited a decrease in the expression of K channels. We then tested the time and dose effects of on the cell viability of INS-1 cells. We observed a significant decrease in the surface expression of K channels after 72 h of treatment with 0.4 mM . In addition, this treatment induced pancreatic β-cell apoptosis by increasing cleaved caspase 3 protein level. Our results demonstrated cotreatment with glibenclamide, the sulfonylurea compounds for type II , in -treated cells reduces cell death and recovers the glucose stimulated insulin secretion through increasing the surface expression of K channels. Importantly, glibenclamide also improved glucose tolerance, triglyceride concentration, and insulin sensitivity in the -injected mice. In conclusion, an increase in the surface expression of K channels restores insulin secretion, reduces pancreatic β-cell\'s apoptosis, highlighting correct trafficking of K channels is important in survival of β-cells during lipotoxicity.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Protective effects of Danzhi jiangtang capsule on vascular endothelial damages induced by high-fat diet and .

Danzhi jiangtang capsule (DJC) is mainly used to treat type 2 and vascular complication. Our aim was to investigate whether the protective effects of DJC by oral administration on high-fat diet (HFD) and -induced damages of endothelial cells are related to oxidative stress and endoplasmic reticulum (ER) stress. Male Sprague-Dawley rats were randomly divided into standard chow diet (SCD), HFD, HFD plus DJC-low dose (HFD\u2009+\u2009DJCL) and HFD plus DJC-high dose treatment groups (HFD\u2009+\u2009DJCH). The latter three groups were given HFD feeding for three months, then the HFD\u2009+\u2009DJCL and HFD\u2009+\u2009DJCH rats were treated with DJC (500 and 1000\u2009mg/kg/day) via gavage for another two months respectively. Endothelium-dependent relaxations induced by acetylcholine were observed in isolated rat thoracic aortic ring preparation. Malondialdehyde (MDA), total-antioxidant capacity (T-AOC), superoxide dismutase (SOD), interleukin 1β (IL-1β), tumour necrosis factor α (TNFα), free fatty acids (FFA), endothelin-1 (ET-1) and nitric oxide (NO) levels in serum were assayed. The pathological changes, protein expression of endothelial NO synthase (eNOS), phosphorylated eNOS (p-eNOS) and ER stress-related genes in the thoracic aorta were measured. Human umbilical vein endothelial cells (HUVEC) were treated with serum-medicated DJC and then with to detect the reactive oxygen species (ROS) levels and C/EBP homologous protein (CHOP) distribution, expression of p-eNOS and ER stress-related genes. DJC therapy exhibited a potent antioxidant activity and effectively prevented the endothelial dysfunction (ED) manifested by promoting p-eNOS expression and enhancing NO release, decreasing lipid deposition (Oil-red O staining, CPT1b and ACC) and inflammation (IL-1β, TNFα, CD68 and p-JNK), alleviating oxidative and ER stress, and decreasing the apoptosis of endothelial cells (TUNEL, BCL-2 and BAX) induced by HFD and respectively. These findings suggest that the administration of DJC had endothelial protective effects against HFD-induced ED, inflammation and apoptosis by alleviating lipid deposition, oxidative and ER stress.Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Keyword: diabetes

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to obesity-related insulin resistance and type 2 (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular energy sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the mitochondria-generated reactive oxygen species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including obesity, insulin resistance, and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: diabetes

Metabolomics Study of Metabolic Changes in Renal Cells in Response to High-Glucose Exposure Based on Liquid or Gas Chromatography Coupled With Mass Spectrometry.

Diabetic nephropathy (DN) is one of the most serious microvascular complications and the leading causes of death in (DM). To find biomarkers for prognosing the occurrence and development of DN has significant clinical value for its prevention, diagnosis, and treatment. In this study, a non-targeted cell metabolomics-based ultra-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry and gas chromatography coupled with mass spectrometry was developed and performed the dynamic metabolic profiles of rat renal cells including renal tubular epithelial cells (NRK-52E) and glomerular mesangial cells (HBZY-1) in response to high glucose at time points of 12\u2009h, 24 h, 36 h, and 48 h. Some potential biomarkers were then verified using clinical plasma samples collected from 55 healthy volunteers, 103 DM patients, and 57 DN patients. Statistical methods, such as principal component analysis and partial least squares to latent structure-discriminant analysis were recruited for data analyses. As a result, and linoleic (all-cis-9,12) were the potential indicators for the occurrence and development of DN, and valine, leucine, and isoleucine could be used as the prospective biomarkers for DM. In addition, rise and fall of leucine and isoleucine levels in plasma could be used for prognosing DN in DM patients. Through this study, we established a novel non-targeted cell dynamic metabolomics platform and identified potential biomarkers that may be applied for the diagnosis and prognosis of DM and DN.

Keyword: diabetes

Hepatocyte growth factor alleviates hepatic insulin resistance and lipid accumulation in high-fat diet-fed mice.

Type 2 is frequently accompanied by fatty liver disease. Lipid accumulation within the liver is considered as one of the risk factors for insulin resistance. Hepatocyte growth factor (HGF) is used to treat liver dysfunction; however, the effect and mechanism of HGF on hepatic lipid metabolism are still not fully understood.Male C57BL/6 mice were induced with a high-fat diet for 12 weeks, followed by a 4-week treatment of HGF or vehicle saline. The levels of fasting blood glucose, fasting insulin and homeostatic model assessment of insulin resistance were calculated for insulin sensitivity. Biochemical plasma parameters were also measured to assess the effect of HGF on lipid accumulation. Additionally, genes in the lipid metabolism pathway were evaluated in -treated HepG2 cells and high-fat diet mice.HGF treatment significantly decreased the levels of fasting blood glucose, hepatic triglyceride and cholesterol contents. Additionally, HGF-regulated expression levels of sterol regulatory element-binding protein-1c/fatty synthase, peroxidase proliferator-activated receptor-α, and upstream nuclear receptors, such as farnesoid X receptor and small heterodimer partner. Furthermore, c-Met inhibitor could partially reverse the effects of HGF.HGF treatment can ameliorate hepatic insulin resistance and steatosis through regulation of lipid metabolism. These effects might occur through farnesoid X receptor-small heterodimer partner axis-dependent transcriptional activity.© 2018 The Authors. Journal of Investigation published by Asian Association for the Study of (AASD) and John Wiley & Sons Australia, Ltd.

Keyword: diabetes

Resveratrol attenuates type 2 by mediating mitochondrial biogenesis and lipid metabolism via Sirtuin type 1.

The rising incidence of type 2 (T2DM) is a major public health problem and novel therapeutic strategies are required to prevent and treat T2DM. It has been demonstrated that resveratrol (RSV) may prevent T2DM by targeting Sirtuin type 1 (SIRT1), indicating that SIRT1 may be a novel therapeutic target for T2DM prevention. In the present study, a T2DM rat model was established by administering a high fat diet and streptozotocin (STZ) injections. Measurements of blood glucose and insulin confirmed successful establishment of the T2DM model. RSV was used to treat rats with STZ-induced T2DM and the results indicated that RSV reversed the STZ-induced downregulation of peroxisome proliferator-activated receptor-γ coactivator-1α, SIRT1 and forkhead box protein O 3a. Furthermore, RSV modulated the activity of superoxide dismutase and malondialdehyde, which are associated with oxidative stress. , cells from the insulinoma cell line clone 1E were pretreated with (PA) to simulate a high fat environment. The results of reverse transcription-quantitative polymerase chain reaction indicated that PA suppressed the expression of SIRT1 in a dose- and time-dependent manner. Furthermore, PA modulated the expression of mitochondrial biogenesis-associated, lipid metabolism-associated and β-cell-associated genes, whereas RSV treatment ameliorated the PA-induced changes in the expression of these genes via SIRT1. The results of the present study suggest that RSV participates in the prevention of T2DM by regulating the expression of mitochondrial genes associated with biogenesis, lipid metabolism and β-cells via SIRT1. The results of the current study provide an insight into the mechanisms by which SIRT1 inhibits T2DM and may be used as a basis for future studies.

Keyword: diabetes

Attenuating effect of silibinin on -induced apoptosis and mitochondrial dysfunction in pancreatic β-cells is mediated by estrogen receptor alpha.

High levels of circulating free fatty acids often trigger pancreatic β cell dysfunction during the development of type 2 . Silibinin, the main component of Silybum marianum fruit extract (silymarin), is reported to have anti-diabetic effect. This study is designed to determine the protective effect of silibinin on -induced damage in a rat pancreatic β-cell line, INS-1 cells. Our results demonstrate that silibinin improves cell viability, enhances insulin synthesis and secretion, and resumes normal mitochondrial function in -treated INS-1 cells. An accumulating body of evidence has shown that the estrogen receptors are key molecules involved in glucose and lipid metabolism. Our results suggest that silibinin upregulates ERα signaling pathway from the finding that ERα-specific inhibitors abolish the anti-lipotoxic effect of silibinin. In conclusion, these findings suggest that silibinin protects INS-1 cells against apoptosis and mitochondrial damage through upregulation of ERα pathway.

Keyword: diabetes

lncRNA MEG3 promotes hepatic insulin resistance by serving as a competing endogenous RNA of miR-214 to regulate ATF4 expression.

MicroRNA (miR)‑214 has been demonstrated to suppress gluconeogenesis by targeting activating transcription factor 4 (ATF4), which regulates gluconeogenesis by affecting the transcriptional activity of forkhead box protein O1 (FoxO1). Our previous study revealed that the upregulation of maternally expressed gene 3 (MEG3), a long noncoding RNA, enhanced hepatic insulin resistance via increased FoxO1 expression. The present study aimed to explore whether miR‑214 and ATF4 were involved in the MEG3‑mediated increase of FoxO1 expression. MEG3, miR‑214 and ATF4 expression were examined by reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among MEG3, miR‑214 and ATF4 was analysed using the luciferase reporter assay. MEG3‑targeting small interference RNAs were injected into high‑fat diet (HFD)‑fed mice to verify the role of MEG3 in hepatic insulin resistance in\xa0vivo. MEG‑3 and ATF4 were demonstrated to be upregulated and miR‑214 was indicated to be downregulated in the livers of HFD‑fed and ob/ob mice. In mouse primary hepatocytes, palmitate time‑dependently increased MEG3 and ATF4 but decreased miR‑214 expression levels. Furthermore, MEG3 served as a competing endogenous RNA (ceRNA) for miR‑214 to facilitate ATF4 expression, while miR‑214 inhibition and ATF4 overexpression reversed the MEG3 knockdown‑mediated decrease in the expression of FoxO1 and FoxO1‑downstream targets phosphoenolpyruvate carboxykinase and glucose‑6‑phosphatase catalytic subunit. In HFD‑fed mice, MEG3 knockdown substantially improved impaired glucose and insulin tolerance, while downregulating HFD‑induced ATF4 expression and upregulating HFD‑suppressed miR‑214 expression. In conclusion, MEG3 promoted hepatic insulin resistance by serving as a ceRNA of miR‑214 to facilitate ATF4 expression. These data provide insight into the molecular mechanism of MEG3 involvement in the development of type 2 .

Keyword: diabetes

Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced insulin resistance in human skeletal muscle cells.

Free fatty is considered to be one of the major pathogenic factors of inducing insulin resistance. The association between iron disturbances and insulin resistance has recently begun to receive a lot of attention. Although skeletal muscles are a major tissue for iron utilization and storage, the role of iron in palmitate (PA)-induced insulin resistance is unknown. We investigated the molecular mechanism underlying iron dysregulation in PA-induced insulin resistance. Interestingly, we found that PA simultaneously increased intracellular iron and induced insulin resistance. The iron chelator deferoxamine dramatically inhibited PA-induced insulin resistance, and iron donors impaired insulin sensitivity by activating JNK. PA up-regulated transferrin receptor 1 (tfR1), an iron uptake protein, which was modulated by iron-responsive element-binding proteins 2. Knockdown of tfR1 and iron-responsive element-binding proteins 2 prevented PA-induced iron uptake and insulin resistance. PA also translocated the tfR1 by stimulating calcium influx, but the calcium chelator, BAPTA-AM, dramatically reduced iron overload by inhibiting tfR1 translocation and ultimately increased insulin sensitivity. Iron overload may play a critical role in PA-induced insulin resistance. Blocking iron overload may thus be a useful strategy for preventing insulin resistance and .-Cui, R., Choi, S.-E., Kim, T. H., Lee, H. J., Lee, S. J., Kang, Y., Jeon, J. Y., Kim, H. J., Lee, K.-W. Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced insulin resistance in human skeletal muscle cells.

Keyword: diabetes

Circulating sphingolipids, fasting glucose, and impaired fasting glucose: The Strong Heart Family Study.

Animal studies suggest sphingolipids as an early marker of impaired glucose metabolism; however, research in humans is limited. We evaluated whether individual sphingolipid species were associated with fasting plasma glucose and incident impaired fasting glucose in a longitudinal cohort study.We measured 15 sphingolipid species from blood samples collected in 2001-2003 from 2145 participants without prevalent in the Strong Heart Family Study. Fasting plasma glucose was measured in blood samples collected at baseline and follow-up (mean 5.5\u202fyears after baseline).The average age of study participants was 38\u202fyears; 41% were men. Ceramide, sphingomyelin, and glucosylceramide species levels were higher in older participants; lactosyl-ceramide levels were higher in participants with lower BMIs. In adjusted analyses, greater concentrations of most ceramide species and lower lactosyl-ceramide with (LC-16) were associated with higher glucose levels at baseline. We did not observe associations of sphingomyelin species or glucosyl-ceramide species with glucose levels. Associations of sphingolipid levels with fasting glucose levels at follow-up were similar but had greater uncertainty than associations with baseline glucose. Although no statistically significant associations of sphingolipids with incident impaired fasting glucose were present, results were similar to glucose analyses.We identified several ceramide species associated with higher fasting glucose levels and one sphingolipid, LC-16, that was associated with lower fasting glucose levels. These findings compliment previous research, which linked these sphingolipids with fasting insulin levels, and suggest that higher levels of these ceramides and lower LC-16 may be an early marker of impaired glucose metabolism. FUND: US National Institutes Health.Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: diabetes

Endogenous Fatty Acids Are Essential Signaling Factors of Pancreatic β-Cells and Insulin Secretion.

The secretion of insulin from β-cells depends on extracellular factors, in particular glucose and other small molecules, some of which act on G-protein-coupled receptors. Fatty acids (FAs) have been discussed as exogenous secretagogues of insulin for decades, especially after the FA receptor GPR40 (G-protein-coupled receptor 40) was discovered. However, the role of FAs as endogenous signaling factors has not been investigated until now. In the present work, we demonstrate that lowering endogenous FA levels in β-cell medium by stringent washing or by the application of FA-free (FAF) BSA immediately reduced glucose-induced oscillations of cytosolic Ca ([Ca] oscillations) in MIN6 cells and mouse primary β-cells, as well as insulin secretion. Mass spectrometry confirmed BSA-mediated removal of FAs, with , stearic, oleic, and elaidic being the most abundant species. [Ca] oscillations in MIN6 cells recovered when BSA was replaced by buffer or as FA levels in the supernatant were restored. This was achieved by recombinant lipase-mediated FA liberation from membrane lipids, by the addition of FA-preloaded FAF-BSA, or by the photolysis of cell-impermeant caged FAs. Our combined data support the hypothesis of FAs as essential endogenous signaling factors for β-cell activity and insulin secretion.© 2018 by the American Association.

Keyword: diabetes

MIP-1α Induction by Palmitate in the Human Monocytic Cells Implicates TLR4 Signaling Mechanism.

MIP-1α (macrophage inflammatory protein 1α)/CCL3 chemokine is associated with the adipose tissue inflammation in obesity. Both MIP-1α and free fatty acids are elevated in obesity/T2D. We asked if free fatty palmitate could modulate MIP1α expression in the human monocytic cells.Human monocytic THP-1 cells and macrophages were stimulated with palmitate and TNF-α (positive control). MIP-1α expression was measured with real time RT-PCR, Flow Cytometry and ELISA. Signaling pathways were identified by using THP-1-XBlue™ cells, THP-1-XBlue™-defMyD cells, anti-TLR4 mAb and TLR4 siRNA.Our data show that palmitate induced significant increase in MIP1α production in monocytic THP-1 cells/macrophages. MIP-1α induction was significantly suppressed when cells were treated with anti-TLR4 antibody prior stimulation with palmitate. Using TLR4 siRNA, we further demonstrate that palmitate-induced MIP-1α expression in monocytic cells requires TLR4. Moreover, THP1 cells defective in MyD88, a major adaptor protein involved in TLR4 signaling, were unable to induce MIP-1α production in response to palmitate. Palmitate-induced MIP-1α expression was suppressed by inhibition of MAPK, NFkB and PI3K signaling pathways. In addition, palmitate-induced NF-κB/AP-1 activation was observed while production of MIP-1α. However, this activation of NF-κB/AP-1 was abrogated in MyD88 deficient cells.Overall, these results show that palmitate induces TLR4dependent MIP-1α expression requiring the MyD88 recruitment and activation of MAPK, NF-κB/AP-1 and PI3K signaling. It implies that the increased systemic levels of free fatty palmitate in obesity/T2D may contribute to metabolic inflammation through excessive production of MIP-1a.© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Keyword: diabetes

-Induced NAD Depletion is Associated with the Reduced Function of SIRT1 and Increased Expression of BACE1 in Hippocampal Neurons.

Increased levels of circulating fatty acids, such as (PA), are associated with the development of obesity, insulin resistance, type-2 and metabolic syndrome. Furthermore, these diseases are linked to an increased risk of cancer, cardiovascular diseases, mild cognitive impairment and even Alzheimer\'s disease (AD). However, the precise actions of elevated PA levels on neurons and their association with neuronal metabolic disruption that leads to the expression of pathological markers of AD, such as the overproduction and accumulation of the amyloid-β peptide, represent an area of intense investigation. A possible molecular mechanism involved in the effects of PA may be through dysfunction of the NAD sensor enzyme, SIRT1. Therefore, the aim of the present study was to analyze the relationship between the effects of PA metabolism on the function of SIRT1 and the upregulation of BACE1 in cultured hippocampal neurons. PA reduced the total amount of NAD in neurons that caused an increase in p65 K310 acetylation due to inhibition of SIRT1 activity and low protein content. Furthermore, BACE1 protein and its activity were increased, and BACE1 was relocated in neurites after PA exposure.

Keyword: diabetes

The effect of FADS2 gene rs174583 polymorphism on desaturase activities, fatty profile, insulin resistance, biochemical indices, and incidence of type 2 .

In this study, we investigated the associations of erythrocytes fatty composition, activities of delta-5 desaturase (D5D) and delta-6 desaturase (D6D), and other metabolic risk factors, with type 2 (T2D) risk to determine if rs174583 polymorphism of FADS2 gene had any effect on these associations.Fatty profile of erythrocytes was determined using gas chromatography-mass spectrometry in 95 T2D patients and 95 apparently healthy participants. The genotypes of single-nucleotide polymorphism (SNP) of FADS2 gene were determined using the polymerase chain reaction-restriction fragment length polymorphism technique. Other biochemical parameters were measured in the serum using standard analytical procedures.D6D activity was increased ( < 0.001) and D5D activity was decreased in T2D patients ( < 0.001) compared to controls. Homeostatic model assessment insulin resistance (HOMA-IR) index was positively correlated with D6D ( = 0.34, < 0.001) and negatively correlated with D5D ( = -0.19, = 0.02). ( < 0.001) and dihomo-gamma-linolenic ( = 0.03) were higher and linoleic ( < 0.001) and arachidonic (AA) ( < 0.001) were lower in T2D patients. The distribution of rs174583 genotypes which includes C/T, C/C, and T/T was not different in the two groups ( = 0.63).In the population studied, there was a strong association in the erythrocytes fatty composition, D5D and D6D activities and other metabolic risk factors between non-T2D and T2D patients. In addition, there was a strong association in erythrocytes DGLA and AA contents and D5D activities between rs174583 genotypes in all participants. However, the distribution of rs174583 genotypes did not differ significantly between T2D patient and controls, and it did not appear to be an association between rs174583 SNP and incident of T2D.

Keyword: diabetes

Induces Müller Cell Inflammation that is Potentiated by Co-treatment with Glucose.

Chronic hyperglycemia is thought to be the major stimulator of retinal dysfunction in diabetic retinopathy (DR). Thus, many -related systemic factors have been overlooked as inducers of DR pathology. Cell culture models of retinal cell types are frequently used to mechanistically study DR, but appropriate stimulators of DR-like factors are difficult to identify. Furthermore, elevated glucose, a gold standard for cell culture treatments, yields little to no response from many primary human retinal cells. Thus, the goal of this project was to demonstrate the effectiveness of the free fatty , and compare its use alone and in combination with elevated glucose as a stimulus for human Müller cells, a retinal glial cell type that is activated early in DR pathogenesis and uniquely responsive to fatty acids. Using RNA sequencing, we identified a variety of DR-relevant pathways, including NFκB signaling and inflammation, intracellular lipid signaling, angiogenesis, and MAPK signaling, that were stimulated by , while elevated glucose alone did not significantly alter any -relevant pathways. Co-treatment of high glucose with potentiated the expression of several DR-relevant angiogenic and inflammatory targets, including PTGS2 (COX-2) and CXCL8 (IL-8).

Keyword: diabetes

Crucial Roles of 5-HT and 5-HT2 Receptor in -Related Lipid Accumulation and Pro-Inflammatory Cytokine Generation in Hepatocytes.

Previously, we confirmed that liver-synthesized 5-HT rather than non-liver 5-HT, acting on the 5-HT2 receptor (5-HT2R), modulates lipid-induced excessive lipid synthesis (ELS). Here, we further revealed the effects of the hepatocellular 5-HT system in -related disorders.Studies were conducted in male ICR mice, human HepG2 cells, and primary mouse hepatocytes (PMHs) under gene or chemical inhibition of the 5-HT system, key lipid metabolism, and inflammation-related factors. Protein and messenger RNA expression and levels of the factors were determined via western blotting, reverse transcription PCR, and quantitative assay kits, respectively. Hepatic steatosis with inflammation and fibrosis, intracellular lipid droplet accumulation (LDA), and reactive oxygen species (ROS) location were determined via hematoxylin and eosin, Masson\'s trichrome, Oil red O, and fluorescent-specific staining, respectively. induced the activation of the 5-HT system: the activation of 5-HT2R, primarily 5-HT2AR, in addition to upregulating monoamine oxidase A (MAO-A) expression and 5-HT synthesis, by activating the G protein/ phospholipase C pathway modulated PKCε activation, resulting in ELS with LDA; the activation of NF-κB, which mediates the generation of pro-inflammatory cytokines, was primarily due to ROS generation in the mitochondria induced by MAO-A-catalyzed 5-HT degradation, and secondarily due to the activation of PKCε. These effects of the 5-HT system were also detected in - or high glucose-treated PMHs and regulated multiple inflammatory signaling pathways. In diabetic mice, co-treatment with antagonists of both 5-HT synthesis and 5-HT2R significantly abolished hepatic steatosis, inflammation, and fibrosis as well as hyperglycemia and dyslipidemia.Activation of the hepatocellular 5-HT system plays a crucial role in inducing -related hepatic dysfunction and is a potential therapeutic target.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: diabetes

Identification of early biological changes in palmitate-treated isolated human islets.

Long-term exposure to elevated levels of free fatty acids (FFAs) is deleterious for beta-cell function and may contribute to development of type 2 (T2DM). Whereas mechanisms of impaired glucose-stimulated insulin secretion (GSIS) in FFA-treated beta-cells have been intensively studied, biological events preceding the secretory failure, when GSIS is accentuated, are poorly investigated. To identify these early events, we performed genome-wide analysis of gene expression in isolated human islets exposed to fatty palmitate for different time periods.Palmitate-treated human islets showed decline in beta-cell function starting from day two. Affymetrix Human Transcriptome Array 2.0 identified 903 differentially expressed genes (DEGs). Mapping of the genes onto pathways using KEGG pathway enrichment analysis predicted four islet biology-related pathways enriched prior but not after the decline of islet function and three pathways enriched both prior and after the decline of islet function. DEGs from these pathways were analyzed at the transcript level. The results propose that in palmitate-treated human islets, at early time points, protective events, including up-regulation of metallothioneins, tRNA synthetases and fatty -metabolising proteins, dominate over deleterious events, including inhibition of fatty detoxification enzymes, which contributes to the enhanced GSIS. After prolonged exposure of islets to palmitate, the protective events are outweighed by the deleterious events, which leads to impaired GSIS.The study identifies temporal order between different cellular events, which either promote or protect from beta-cell failure. The sequence of these events should be considered when developing strategies for prevention and treatment of the disease.

Keyword: diabetes

Oxidative stress induced by modulates K2.3 channels in vascular endothelium.

Elevated plasma free fatty acids level has been implicated in the development of insulin resistance, inflammation, and endothelial dysfunction in diabetic and nondiabetic individuals. However, the underlying mechanisms still remain to be defined. Herein, we investigated the effect of (PA), the most abundant saturated fatty in the human body, on small-conductance Ca-activated potassium channels (K2.3)-mediated relaxation in rodent resistance arteries and the underlying molecular mechanism. The effect of PA on K2.3 in endothelium was evaluated using real-time PCR, Western blotting, whole-cell patch voltage-clamp, wire and pressure myograph system, and reactive oxygen species (ROS) were measured by using dihydroethidium and 2\', 7\'-dichlorofluorescein diacetate. K2.3-mediated vasodilatation responses to acetylcholine and NS309 (agonist of K2.3 and K3.1) were impaired by incubation of normal mesenteric arteries with 100\u202fμM\u202fPA for 24\u202fh. In cultured human umbilical vein endothelial cells (HUVECs), PA decreased K2.3 current and expression at mRNA and protein levels. Incubation with the NADPH oxidase (Nox) inhibitor dibenziodolium (DPI) partly inhibited the PA-induced ROS production and restored K2.3 expression. Inhibition of either p38-MAPK or NF-κB using specific inhibitors (SB203580, SB202190 or Bay11-7082, pyrrolidinedithiocarbamate) attenuated PA-induced downregulation of K2.3 and inhibition of p38-MAPK also attenuated PA-induced phosphorylation of NF-κB p65. Furthermore, DPI reversed the increment of phospho-p38-MAPK by PA. These results demonstrated that PA downregulated K2.3 expressions via Nox/ROS/p38-MAPK/NF-κB signaling leading to endothelial vasodilatory dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: diabetes

Acclimation of CC myoblasts to physiological glucose concentrations for in vitro research.

The interplay between hyper-glycemia and -lipidemia in (DM) is important in simulating diabetic conditions. However, cell culture media typically contain supraphysiological levels of glucose to stimulate cellular growth, which also desensitizes cells to elevated glucose levels. Moreover, creating hyperlipidemic conditions in vitro requires specialized carriers because unbound lipids form micelles when introduced to liquid media. This study sought to develop a novel method for simulating DM conditions in vitro.We acclimated the CC mouse myoblasts to culture medium with 5.6\u202fmM glucose, which mimics physiological levels, and created a bovine serum albumin- conjugate for lipid transport to explore the effects of hyperlipidemia. We simulated diabetic conditions in vitro by using both hyper-glycemic and -lipidemic conditions and compared the results to that of only hyperglycemic or hyperlipidemic conditions.Acclimated cells exposed to these hyper-glycemic (15\u202fmM glucose) and/or -lipidemic (0.25\u202fmM palmitate) conditions for 2\u202fh showed increased mitochondrial fragmentation and membrane potential as well as elevated reactive oxygen species production compared to control cells. These findings suggest altered mitochondrial morphology and function, which have been confirmed using isolated rat flexor digitorum brevis myofibers. Hyper-glycemic and/or -lipidemic stimulations for 24\u202fh significantly increased mitogen-activated protein kinase kinase MEK 1/2 protein expression, upregulated the early pro-apoptotic transcription factor C/EBP homologous protein (CHOP), and induced apoptosis.Our results further support and confirm the utility of this method which will allow for subsequent investigations studying the effects of hyper-glycemia and/or -lipidemia in vitro.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Adiponectin homolog novel osmotin protects obesity/-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic fatty liver disease (NAFLD) in obesity-associated .To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human liver carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum lipid parameters, and increased fatty oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: diabetes

Time-Dependent alteration to the tight junction structure of distal intestinal epithelia in type 2 prediabetic mice.

High-fat diet (HFD) intake has been associated with changes in intestinal microbiota composition, increased intestinal permeability, and onset of type 2 (T2DM). The aim of this work was twofold: 1) to investigate the structural and functional alterations of the tight junction (TJ)-mediated intestinal epithelial barrier of ileum and colon, that concentrate most of the microbiota, after exposure to a HFD for 15, 30 and 60 days, and 2) to assess the effect of in vitro exposure to free fatty acids (FFAs), one of the components of HFD, on paracellular barrier of colon-derived Caco-2\u202fcells.HFD exposure induced progressive metabolic changes in male mice that culminated in prediabetes after 60d. Morphological analysis of ileum and colon mucosa showed no signs of epithelial rupture or local inflammation but changes in the junctional content/distribution and/or cellular content of TJ-associated proteins (claudins-1, -2, -3, and occludin) in intestinal epithelia were seen mainly after a prediabetes state has been established. This impairment in TJ structure was not associated with significant changes in intestinal permeability to FITC-dextran. Exposure of Caco-2 monolayers to or linoleic acids seems to induce a reinforcement of TJ structure while treatment with oleic had a more diverse effect on TJ protein distribution.TJ structure in distal intestinal epithelia can be specifically impaired by HFD intake at early stage of T2DM, but not by FFAs in vitro. Since the TJ change in ileum/colon was marginal, probably it does not contribute to the disease onset.Copyright © 2019. Published by Elsevier Inc.

Keyword: diabetes

Epigallocatechin gallate improves insulin resistance in HepG2 cells through alleviating inflammation and lipotoxicity.

High levels of circulating free fatty acids (FFAs), inflammation and oxidative stress are important causes for insulin resistance (IR) and type 2 . The aim of this study was to investigate the mechanisms of EGCG in alleviating IR in HepG2 cells.HepG2 cells were treated with 25\u202fmM glucose, 0.25\u202fmM (PA), or 50\u202fμM EGCG for 24\u202fh.EGCG increased glucose uptake and decreased glucose content. EGCG markedly decreased the levels of inflammatory and oxidative stress factors including nuclear factor κB (NF-κB), tumor necrosis factor-α, interleukin-6, reactive oxygen species, malondialdehyde and p53 protein, and markedly increased superoxide dismutases (SOD), glutathione peroxidase and SOD2 protein. EGCG significantly downregulated the levels of FFAs, triacylglycerol and cholesterol in HepG2 cells. The glucose transporter 2 (GLUT2) protein and its downstream proteins peroxisome proliferator-activated receptor γ coactivator (PGC)-1β were significantly increased, and sterol regulatory element-binding-1c (SREBP-1c) protein, and fatty synthase (FAS) were significantly decreased by EGCG in HepG2. Moreover, the foregoing effects were reversed by siRNA-mediated knockdown of GLUT2.Our data demonstrated that EGCG improved IR, possibly through ameliorating glucose (25\u202fmM) and PA (0.25\u202fmM)-induced inflammation, oxidative stress, and FFAs via the GLUT2/PGC-1β/SREBP-1c/FAS pathway in HepG2 cells.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

A novel PPARα/γ agonist, propane-2-sulfonic octadec-9-enyl-amide, ameliorates insulin resistance and gluconeogenesis in vivo and vitro.

Peroxisome proliferator-activated receptor alpha/gamma (PPARα/γ) agonists have emerged as important pharmacological agents for improving insulin action. Propane-2-sulfonic octadec-9-enyl-amide (N15) is a novel PPARα/γ dual agonist synthesized in our laboratory. The present study investigates the efficacy and safety of N15 on insulin resistance regulation in high fat diet (HFD)-and streptozotocin (STZ)-induced diabetic mice and in (PA)-induced HepG2 cells. Our results showed that N15 remarkably ameliorated insulin resistance and dyslipidemia in vivo, as well as rectified the glucose consumption and gluconeogenesis in vitro. Moreover, the glucose-lowering effect of N15 was associated with PPARγ mediated up-regulation of hepatic glucose consumption and down-regulation of gluconeogenesis. Meanwhile, N15 exerted advantageous effects on glucose and lipid metabolism without triggering weight gain and hepatotoxicity in mice. In conclusion, our data demonstrated that by alleviating glucose and lipid abnormalities, N15 could be used as a potential prophylactic and therapeutic agent against type 2 and related metabolic disorders.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: diabetes

Fatty and Lipopolysaccharide Effect on Beta Cells Proteostasis and its Impact on Insulin Secretion.

Metabolic overload by saturated fatty acids (SFA), which comprises β-cell function, and impaired glucose-stimulated insulin secretion are frequently observed in patients suffering from obesity and type 2 . The increase of intracellular Ca triggers insulin granule release, therefore several mechanisms regulate Ca efflux within the β-cells, among others, the plasma membrane Ca-ATPase (PMCA). In this work, we describe that lipotoxicity mediated mainly by the saturated (PA) (16C) is associated with loss of protein homeostasis (proteostasis) and potentially cell viability, a phenomenon that was induced to a lesser extent by stearic (18C), myristic (14C) and lauric (12C) acids. PA was localized on endoplasmic reticulum, activating arms of the unfolded protein response (UPR), as also promoted by lipopolysaccharides (LPS)-endotoxins. In particular, our findings demonstrate an alteration in PMCA1/4 expression caused by PA and LPS which trigger the UPR, affecting not only insulin release and contributing to β-cell mass reduction, but also increasing reactive nitrogen species. Nonetheless, stearic (SA) did not show these effects. Remarkably, the proteolytic degradation of PMCA1/4 prompted by PA and LPS was avoided by the action of monounsaturated fatty acids such as oleic and palmitoleic . Oleic recovered cell viability after treatment with PA/LPS and, more interestingly, relieved endoplasmic reticulum (ER) stress. While palmitoleic improved the insulin release, this fatty seems to have more relevant effects upon the expression of regulatory pumps of intracellular Ca. Therefore, chain length and unsaturation of fatty acids are determinant cues in proteostasis of β-cells and, consequently, on the regulation of calcium and insulin secretion.

Keyword: diabetes

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic fatty liver disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic fatty liver disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for gluconeogenesis (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in liver. Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for gluconeogenesis in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and lipid metabolism through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose metabolism via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: diabetes

Palmitate and oleate co-treatment increases myocellular protein content via impaired protein degradation.

Protein balance is a crucial determinant of myocellular size and function. The effects of fatty acids on myocellular protein balance remain controversial. The aim of this study was to determine the direct effects of a mixed-species fatty environment on myocellular protein synthesis and degradation.C2C12 myotubes were cultured in media containing equimolar (250\xa0μM) and oleate (PO) or bovine serum albumin control for ≤72\xa0h. Myocellular protein balance was determined via incorporation (synthesis) or release (degradation) of H-tyrosine after 24, 48, and 72\xa0h of treatment. Expression of major proteolytic genes was measured by reverse transcription polymerase chain reaction.PO significantly increased myocellular protein content at 24, 48, and 72\xa0h. Basal myocellular protein synthesis was unchanged by PO. However, PO significantly decreased basal rate of protein degradation at 24\xa0h and this effect persisted throughout 72\xa0h of treatment. Expression of the proteolytic genes Atrogin-1 (MAFbx), MuRF-1, LC3, and ATG4 B, was reduced during the 72\xa0h PO.A mixed-species fatty environment increases myocellular protein content by decreasing the rate of protein degradation, which may be regulated at the level of gene transcription.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: diabetes

, but not high-glucose, induced myocardial apoptosis is alleviated by N‑acetylcysteine due to attenuated mitochondrial-derived ROS accumulation-induced endoplasmic reticulum stress.

Pharmacological inhibition of reactive oxygen species (ROS) is a potential strategy to prevent -induced cardiac dysfunction. This study was designed to investigate precise effects of antioxidant N‑acetylcysteine (NAC) in alleviating diabetic cardiomyopathy (DCM). Echocardiography and histologic studies were performed 12 weeks after streptozocin injection. Protein levels involved in endoplasmic reticulum stress (ERS) and apoptosis were analyzed by western blotting in diabetic hearts or high-glucose (HG, 30\u2009mM)- and (PA, 300\u2009μM)-cultured neonatal rat cardiomyocytes (NRCMs). ROS generation and structural alterations of mitochondria were also assessed. We report that NAC alleviated -induced cardiac abnormality, including restored ejection fraction (EF %), fraction shortening (FS %), peak E to peak A ratio (E/A) and reduced cardiac hypertrophy and fibrosis. These effects were concomitant with blocked ERS and apoptosis, as evidenced by inactivation of phosphorylated inositol-requiring enzyme-1α (IRE1α)/spliced X-box binding protein 1 (XBP1), phosphorylated protein kinase-like kinase (PERK)/phosphorylated eukaryotic initiation factor 2α (eIF2α) and glucose-regulated protein 78 (GRP78)/activating transcription factor 6 (ATF6α)/C/EBP homologous protein (CHOP) pathways, as well as suppressed Bcl-2-associated X protein (BAX)/B-cell lymphoma-2 (Bcl-2) and cleaved caspase 3 expressions. Mechanistically, PA mediated excessive mitochondrial ROS generation and oxidative stress, which were antagonized by NAC and Mito-TEMPO, a mitochondrial ROS inhibitor. No effects were noted by addition of apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, and NADPH oxidase 4 (NOX 4) and NOX 2 expressions were not altered, indicating that PA-induced ROS generation is independent of NADPH oxidases. Most intriguingly, HG failed to promote ROS production despite its ability to promote ERS and apoptosis in NRCMs. Collectively, these findings indicate that NAC primarily abrogates PA-mediated mitochondrial ROS through ERS and therefore alleviates myocardial apoptosis but has little effect on HG-induced cardiac injury. This uncovers a potential role for NAC in formulating novel cardioprotective strategies in DCM patients.

Keyword: diabetes

The novel NADPH oxidase 4 selective inhibitor GLX7013114 counteracts human islet cell death in vitro.

It has been proposed that pancreatic beta-cell dysfunction in type 2 is promoted by oxidative stress caused by NADPH oxidase (Nox) over-activity. The aim of the present study was to evaluate the efficacy of novel Nox inhibitors as protective agents against cytokine- or high glucose + palmitate-induced human beta-cell death. The Nox2 protein was present mainly in the cytoplasm and was induced by cytokines. Nox4 protein immunoreactivity, with some nuclear accumulation, was observed in human islet cells, and was not affected by islet culture in the presence of cytokines or high glucose + palmitate. Nox inhibitors with partial or no isoform selectivity (DPI, dapsone, GLX351322, and GLX481372) all reduced ROS production of human islet cells exposed to high glucose + palmitate. This was paralleled by improved viability and reduced caspase 3 activation. The Nox1 selective inhibitor ML171 failed to reduce human islet cell death in response to both cytokines and high glucose + palmitate. The selective Nox2 inhibitor Phox-I2 also failed to protect against cytokines, but protected partially against high glucose + palmitate-induced cellular death. The highly selective Nox4 inhibitor GLX7013114 protected islet cells against both cytokines and high glucose + palmitate. However, as no osmotic control for high glucose was used, we cannot exclude the possibility that the high glucose effect was due to osmosis. It is concluded that Nox4 may participate in stress-induced islet cell death in human islets in vitro. We propose that Nox4 mediates pro-apoptotic effects in intact islets under stressful conditions and that selective Nox4-inhibition may be a therapeutic strategy in type 2 .

Keyword: diabetes

Novel Mechanisms Modulating Palmitate-Induced Inflammatory Factors in Hypertrophied 3T3-L1 Adipocytes by AMPK.

A growing body of evidence indicates that AMP-activated protein kinase (AMPK) contributes to not only energy metabolic homeostasis but also the inhibition of inflammatory responses. However, the underlying mechanisms remain unclear. To elucidate the role of AMPK, in this study, we observed the effects of AMPK activation on monocyte chemoattractant protein-1 (MCP-1) release in mature 3T3-L1 adipocytes.We observed signal transduction pathways regulating MCP-1, which increased in obese adipocytes, in an model of hypertrophied 3T3-L1 adipocytes preloaded with palmitate.Palmitate-preloaded cells exhibited significant increase in MCP-1 release and triglyceride (TG) deposition. Increased MCP-1 release and TG deposition were significantly decreased by an AMPK activator. In addition, the AMPK activator not only markedly diminished MCP-1 secretion but also augmented phosphorylation of nuclear factor-B (NF-B) and extracellular signal-regulated kinase (ERK) 1/2. In contrast, MCP-1 release suppression was abolished by the AMPK inhibitor compound C and the MEK inhibitor U0126.MCP-1 release from hypertrophied adipocytes is suppressed by AMPK activation through the NF-B and ERK pathways. These findings provide evidence that AMPK plays a crucial role in ameliorating obesity-induced inflammation.

Keyword: diabetes

Restoration of GLP-1 secretion by Berberine is associated with protection of colon enterocytes from mitochondrial overheating in diet-induced obese mice.

L-cell dysfunction is reported for GLP-1 reduction in type 2 diabetes. However, the mechanism of dysfunction remains unknown. In this study, we examined mitochondrial function in the mechanistic study in diet-induced obese (DIO) mice.C57BL/6 mice were fed a high-fat diet (HFD) for 16 weeks to establish the DIO model for GLP-1 reduction. The mice were then treated with berberine (BBR) (100\u2009mg/kg/day) for 8 weeks to test the impact on GLP-1 expression. Mitochondrial activities of the colon enterocytes were compared among three groups of mice (lean, DIO, and DIO\u2009+\u2009BBR) at the end of treatment. Gut and short-chain fatty acids (SCFAs) were examined to understand the mitochondrial responses. A cellular model treated with (PA) was used in the mechanism study.A reduction in GLP-1 expression was observed in DIO mice with mitochondrial stress responses in the colon enterocytes. The mitochondria exhibited cristae loss, membrane rupture, and mitochondrial swelling, which was observed with an increase in ATP abundance, complex I activity, and deficiency in the activities of complexes II and IV. Those changes were associated with dysbiosis and a reduction in SCFAs in the colon of DIO mice. In the cellular model, an increase in ATP abundance, loss of mitochondrial potential, and elevation of apoptosis were induced by PA. All of the alterations in DIO mice and the cellular model were attenuated by BBR.The mitochondrial stress responses were observed in the colon enterocytes of DIO mice for GLP-1 reduction. The stress was prevented by BBR in the restoration of GLP-1 expression, in which BBR may act through direct and indirect mechanisms.

Keyword: dysbiosis

Gut and Metabolome Response of Seed Oil on Metabolism Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced metabolism disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and lipid metabolism in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan metabolism.

Keyword: dysbiosis

Fatty Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of fatty acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated fatty acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated fatty acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut , and hepatic metabolism.

Keyword: dysbiosis

Vaginal lipidomics of women with vulvovaginal candidiasis and cytolytic vaginosis: A non-targeted LC-MS pilot study.

To characterize the lipid profile in vaginal discharge of women with vulvovaginal candidiasis, cytolytic vaginosis, or no vaginal infection or dysbiosis.Cross-sectional study.Genital Infections Ambulatory, Department of Tocogynecology, University of Campinas, Campinas, São Paulo-Brazil.Twenty-four women were included in this study: eight with vulvovaginal candidiasis, eight with cytolytic vaginosis and eight with no vaginal infections or dysbiosis (control group).The lipid profile in vaginal discharge of the different study groups was determined by liquid chromatography-mass spectrometry and further analyzed with MetaboAnalyst 3.0 platform.Vaginal lipids concentration and its correlation with vulvovaginal candidiasis and cytolytic vaginosis.PCA, PLS-DA and hierarchical clustering analyses indicated 38 potential lipid biomarkers for the different groups, correlating with oxidative stress, inflammation, apoptosis and integrity of the vaginal epithelial tissue. Among these, greater concentrations were found for Glycochenodeoxycholic -7-sulfate, O-adipoylcarnitine, 1-eicosyl-2-heptadecanoyl-glycero-3-phosphoserine, undecanoic , formyl dodecanoate and lipoic in the vulvovaginal candidiasis group; N-(tetradecanoyl)-sphinganine, DL-PPMP, 1-oleoyl-cyclic phosphatidic, and 5-aminopentanoic in the cytolytic vaginosis group; and 1-nonadecanoyl-glycero-3-phosphate, eicosadienoic , 1-stearoyl-cyclic-phosphatidic , 1-(9Z,12Z-heptadecadienoyl)-glycero-3-phosphate, formyl 9Z-tetradecenoate and 7Z,10Z-hexadecadienoic in the control group.Lipids related to oxidative stress and apoptosis were found in higher concentrations in women with vulvovaginal candidiasis and cytolytic vaginosis, while lipids related to epithelial tissue integrity were more pronounced in the control group. Furthermore, in women with cytolytic vaginosis, we observed higher concentrations of lipids related to bacterial overgrowth.

Keyword: dysbiosis

Dynamic alterations in the gut and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). Liver injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with liver fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut and metabolome.

Keyword: dysbiosis

Effects of eicosapentaenoic and gamma-linolenic acids (dietary lipids) on pulmonary surfactant composition and function during porcine .

To investigate whether a diet enriched with fish and borage oils, with their high polyunsaturated fatty (PUFA) content, alters surfactant composition and function during .Prospective, randomized, blinded, controlled animal study.Research laboratory at a medical center.Thirty-six 15- to 25-kg, disease-free, castrated male pigs. DIETS AND MEASUREMENTS: Three groups of pigs (n = 12 per group) were fed for 8 days diets containing either omega-6 fatty acids (FAs) (corn oil; diet A), or omega-3 FAs (fish oil; diet B), or a combination of omega-6 and omega-3 FAs (borage and fish oils; diet C). Eight of 12 pigs in each group received a 0.1-mg/kg bolus of Escherichia coli endotoxin followed by a continuous infusion (0. 075 mg/kg/h). One lung was subsequently isolated ex vivo, and pressure-volume curves were measured. The contralateral lung was lavaged, and surfactant was analyzed for total and individual phospholipids and FA composition. Minimum and maximum surface tension was measured by bubble surfactometry.Pigs fed either diet B or C had increased oleic (C(18:1) omega-9), eicosapentaenoic (EPA; C(20:5) omega-3), docosahexaenoic (C(22:6) omega-3), and total omega-3 and monounsaturated FAs in their surfactant PUFA pools. The relative percentage of linoleic (C(18:2) omega-6) and total omega-6 FAs were significantly lower from pigs fed diets B and C compared with diet A. (C(16:0)) concentrations, the primary FA in surfactant, had a tendency to be lower in pigs fed diets B and C. There were no demonstrable effects on surfactant function or pulmonary compliance.Diets containing EPA or EPA and gamma-linolenic altered the PUFA composition of pulmonary surfactant, but without demonstrable effects on surfactant function during porcine .

Keyword: endotoximia

Deletion of apoptosis signal-regulating kinase 1 (ASK1) protects pancreatic beta-cells from stress-induced death but not from glucose homeostasis alterations under pro-inflammatory conditions.

Type 2 diabetes is characterized by pancreatic beta-cell dysfunction and is associated with low-grade . Recent observations suggest that apoptosis signal-regulating kinase 1 (ASK1) is involved in beta-cell death in response to different stressors. In this study, we tested whether ASK1 deficiency protects beta-cells from glucolipotoxic conditions and cytokines treatment or from glucose homeostasis alteration induced by endotoxemia.Insulin secretion was neither affected upon shRNA-mediated downregulation of ASK1 in MIN6 cells nor in islets from ASK1-deficient mice. ASK1 silencing in MIN6 cells and deletion in islets did not prevent the deleterious effect of glucolipotoxic conditions or cytokines on insulin secretion. However, it protected MIN6 cells from death induced by ER stress or palmitate and islets from short term caspase activation in response to cytokines. Moreover, endotoxemia induced by LPS infusion increased insulin secretion during hyperglycemic clamps but the response was similar in wild-type and ASK1-deficient mice. Finally, insulin sensitivity in the presence of LPS was not affected by ASK1-deficiency.Our study demonstrates that ASK1 is not involved in beta-cell function and dysfunction but controls stress-induced beta-cell death.

Keyword: endotoximia

Effects of endotoxin infusion on circulating levels of eicosanoids, progesterone, cortisol, glucose and lactic , and abortion in pregnant cows.

The effects of Escherichia coli endotoxin infusions (1.0 or 2.5 micrograms kg-1 over 6 h) on pregnancy were investigated in cows in the first, second and third trimester of gestation. Endotoxin increased the plasma levels of prostaglandins (PGs), thromboxane B2 and cortisol, and decreased progesterone. The severity of the clinical signs and the magnitude of the increases in plasma PGs, thromboxane B2 and cortisol tended to depend on the dose of endotoxin, but were independent of the gestation period. There was hyperglycemia followed by hypoglycemia and lactic acidemia. Hyperglycemia and lactic acidemia were significant only at the high dose of endotoxin. Endotoxin infusion at both doses caused a preferential mobilization of oleic from adipose tissue, and also had some effects on the mobilization of and stearic acids during the post-infusion period. The cows in the first trimester of gestation were more sensitive to the abortifacient effect of endotoxin than cows in the second and third trimester of gestation. The results of this study indicate that the mechanism of endotoxin-induced abortion in cows initially involves a prolonged release of PGF2 alpha and its subsequent stimulant effect on uterine smooth muscle contraction and luteolytic effect leading to a gradual decline in the plasma levels of progesterone. It was concluded that pregnancy terminates in the absence of an adequate level of progesterone, especially during the first trimester of gestation, when progesterone of extraluteal origin is not yet available, coupled with the PGF2 alpha-induced propulsive contraction of the uterus. In addition, the metabolic and circulatory failures in severe cases of , especially at the high dose of endotoxin, resulting either directly or indirectly via the release of various autacoids, catecholamines and cortisol, may also contribute to the termination of pregnancy at any stage of gestation.

Keyword: endotoximia

Potential therapeutic efficacy of inhibitors of human phospholipase A2 in septic shock.

Soluble phospholipase A2 has been implicated in the pathogenesis of local and systemic inflammatory reactions. Elevated levels of circulating phospholipase A2 (PLA2) correlate with the severity of circulatory collapse and pulmonary dysfunction in gram-negative septic shock. Characterization of septic shock serum PLA2 revealed a calcium-dependent enzyme with absolute 2-acyl specificity with a pH optimum of 7.5. We tested a number of therapeutic agents for their ability to inhibit PLA2 from human septic shock serum. Chloroquine, chlorpromazine, dexamethasone base, dexamethasone sodium phosphate, indomethacin, lidocaine, oleic , , promethazine, trans-retinoic , rutin and dl-alpha-tocopherol were all studied over the range of 10(-2) to 10(-7) M. All agents, with the sole exception of dexamethasone base, inhibited PLA2 activity at concentrations greater than 10(-3) M. PLA2 inhibition by dexamethasone sodium phosphate was factitious, due to the formation of calcium-phosphate complexes. Of the 11 agents studied, chlorpromazine was the most effective, with an IC50 of 7.5 X 10(-5) M, a membrane concentration achievable within its therapeutic range. Inhibition was non-competitive with an apparent Ki of 5 nM. Since serum PLA2 levels correlate with mortality in both experimental and clinical gram-negative septic shock, and chlorpromazine was previously shown to improve survival in these conditions, we postulate that its therapeutic efficacy resides at least in part in its PLA2-inhibitory activity.

Keyword: endotoximia

[Synthesis of 4-palmitoyl-sinomenine and its anti- activity].

To synthesize the derivatives of sinomenine, 4-palmitoyl-sinomenine, and explore its therapeutic effect on lipopolysaccharide (LPS)-induced endotoxemia.A highly efficient synthesis of sinomenine derivatives called 4-palmitoyl-sinomenine was made with a molecule of substitutions at C-16 position of ring A. One hour before endotoxemia induction by i.p. injection of 10 mg/kg LPS, high-dose treatment mice (n=5/group) received an i.p. injection of 5 mg/kg sinomenine or 4-palmitoyl-sinomenine while the low-dose treatment mice (n=5/group) received 2.5 mg/kg sinomenine or 4-palmitoyl-sinomenine. Untreated group and normal control group received normal saline. And their survival was monitored hourly for 24 hours. Examination of cytotoxicity of 4-palmitoyl-sinomenine on RAW264.7 cells was conducted at a concentration range of 1 to 125 μmol/L using MTT assay. RAW264.7 cells were exposed to 4-palmitoyl-sinomenine (0, 1, 2, 5, 10 μmol/L) for 24 hours, and then treated with LPS (1 μg/mL) for 6 hours. Then RAW264.7 cells were collected and the mRNA level of IL-6 was detected by real-time quantitative PCR(qRT-PCR).Sinomenine derivatives were successfully synthesized to get 4-palmitoyl-sinomenine. The survival percentage of 4-palmitoyl-sinomenine treatment groups was higher than that of sinomenine treatment groups at the same treatment concentration. The 4-palmitoyl-sinomenine inhibited RAW264.7 cell proliferation and IL-6 gene transcription.The 4-palmitoyl-sinomenine has an anti- probably through inhibiting the proliferation of RAW264.7 cells and decreasing the inflammatory gene expression and inflammatory cytokine release.

Keyword: endotoximia

Effects of smooth and rough Pasteurella haemolytica lipopolysaccharides on plasma cyclic-nucleotides and free fatty acids in calves.

The present study examined the potency of smooth or rough Pasteurella haemolytica lipopolysaccharide infusion (LPS, 24 ng kg-1 min-1 for 500 min) on plasma cyclic-nucleotides and several free fatty acids (FFA) in calves. Both smooth or rough LPS increased plasma cAMP immediately to its maximum at 1 h of infusion, whereas plasma cGMP levels rose slowly and peaked 12 h later. The increases in cAMP levels were more prolonged for smooth LPS than rough LPS. The maximum plasma cAMP rise coincided with increases of several plasma FFA. Rough LPS increased plasma oleic greater than greater than stearic greater than linoleic acids in the second hour and reached their steady state levels between 2 h of infusion and 5 h post-infusion. Thereafter, oleic remained maximally elevated, while stearic decreased and other FFA returned to baseline. Smooth LPS had no effects on and stearic acids, but elevated oleic in an essentially similar manner to rough LPS and increased linoleic initially at 5 h, followed by decreases throughout post-infusion. These results demonstrate that produces early marked elevations in plasma cAMP, a gradual rise in plasma cGMP and disproportionate increases in several plasma FFA. The data also demonstrate that smooth and rough LPS differ in their abilities to increase plasma cAMP and FFA and these may be attributed to differences in their in vivo mechanisms of action. The study suggests that cAMP and cGMP may mediate actions of endotoxin at the cellular level and that differences exist in release and/or utilization of each FFA at different stages of .

Keyword: endotoximia

Anti-inflammatory activity of methyl palmitate and ethyl palmitate in different experimental rat models.

Methyl palmitate (MP) and ethyl palmitate (EP) are naturally occurring fatty esters reported as inflammatory cell inhibitors. In the current study, the potential anti-inflammatory activity of MP and EP was evaluated in different experimental rat models. Results showed that MP and EP caused reduction of carrageenan-induced rat paw edema in addition to diminishing prostaglandin E2 (PGE2) level in the inflammatory exudates. In lipopolysaccharide (LPS)-induced in rats, MP and EP reduced plasma levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). MP and EP decreased NF-κB expression in liver and lung tissues and ameliorated histopathological changes caused by LPS. Topical application of MP and EP reduced ear edema induced by croton oil in rats. In the same animal model, MP and EP reduced neutrophil infiltration, as indicated by decreased myeloperoxidase (MPO) activity. In conclusion, this study demonstrates the effectiveness of MP and EP in combating inflammation in several experimental models.Copyright © 2012 Elsevier Inc. All rights reserved.

Keyword: endotoximia

In vitro Modulation of the LPS-Induced Proinflammatory Profile of Hepatocytes and Macrophages- Approaches for Intervention in Obesity?

Low grade endotoxemia is a feature of obesity which is linked to development of steatohepatitis in non-alcoholic fatty liver disease. In this study, macrophages (J774) and hepatocytes (HepG2) were stimulated with lipopolysaccharide (LPS) from E. coli 0111: B4 and analyzed for modulation of this response when preconditioned or stimulated subsequent to LPS, with different doses of Vitamin D3 or docosahexaenoic (DHA) over a time period of 1 and 5 days. Pro-inflammatory TNFα and pro-fibrotic TGFβ released into the supernatants were measured by ELISA; qPCR was performed for Srebp-1c and PPARα mRNA (genes for products involved in fatty synthesis and catabolism, respectively). Vitamin D3 and DHA exerted a consistent, dose dependent anti-inflammatory effect, and increased PPARα relative to Srebp-1c in both cell types. By contrast, addition of free fatty acids (FFA, oleic / 2:1) caused aggravation of LPS-induced inflammatory reaction and an increase of Srebp-1c relative to PPARα. Our results argue in favor of dietary supplementation of Vitamin D3 or DHA (and avoidance of monounsaturated/saturated fatty acids) to alleviate development of fatty liver disease.

Keyword: endotoximia

Saturated fatty combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated fatty (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic , a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Keyword: endotoximia

Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide.

We have previously reported that Smad6, one of the inhibitory Smads of transforming growth factor-β (TGF-β)/bone morphogenetic protein (BMP) signaling, inhibits Toll-like receptor (TLR) 4 signaling by disrupting the Pellino-1-mediated TLR4 signaling complex. Here, we developed Smaducin-6, a novel membrane-tethered -conjugated Smad6-derived peptide composed of amino acids 422-441 of Smad6. Smaducin-6 interacted with Pellino-1, located in the inner membrane, thereby disrupting the formation of IRAK1-, RIP1-, IKKε-mediated TLR4 signaling complexes. Systemic administration of Smaducin-6 showed a significant therapeutic effect on mouse TLR4-mediated inflammatory disease models, cecal-ligation-puncture (CLP)-induced sepsis, and lipopolysaccharide-induced endotoxemia, by inhibiting pro-inflammatory cytokine production and apoptosis while enhancing neutrophil migration and bacterial clearance. Our findings provide clues to develop new peptide-based drugs to target Pellino-1 protein in TLR4 signaling pathway for the treatment of sepsis.© 2015 The Authors. Published under the terms of the CC BY 4.0 license.

Keyword: endotoximia

An insight into nutritional profile of selected Pleurotus species.

The global demand for good quality food indicates that consumers are more concerned about a particular diet associated with good health and lower risk for certain ailments. Mushrooms are widely used as healthy nutritious food. In the present study, the nutritional composition of four different Pleurotus sp. was determined. Prior to extraction, all the selected mushrooms were subjected for proximate composition analysis. The protein, fat, ash, total carbohydrate, fiber and contents were in the range of (16.07- 25.15%), (0.64-2.02%), (2.1-9.14%), (65.66-82.47%), (6.21-54.12%) and (342.20-394.30Kcal/100g), respectively. The spectrophotometric analysis showed that the concentration of protein was in the range of 45.78-33.47 mg/g in all Pleurotus sp. High performance liquid chromatographic analysis of sugars showed six different mono and disaccharides in all the selected mushrooms. The fatty profile by gas chromatography-mass spectrometry revealed that the main fatty acids in selected mushrooms were present in the order linoliec > oleic > . The results suggested that all the Pleurotus sp. could be considered as a rich source of nutrients.

Keyword: energy

Long-term supplementation interacts with parity in lactating dairy cows: Production responses, nutrient digestibility, and partitioning.

The objective of our study was to evaluate the effects of long-term (C16:0) supplementation and parity on production, nutrient digestibility, and partitioning of mid-lactation dairy cows. Forty mid-lactation Holstein cows (18 primiparous and 22 multiparous) were used in a block design. Cows were assigned to receive either a control diet containing no supplemental fat (CON) or a C16:0-enriched supplemented diet (PA; 1.5% diet dry matter) fed for 10 wk. Compared with CON, PA increased dry matter intake, milk yield, cumulative milk yield, milk fat content, milk fat yield, 16-carbon milk fatty (FA) yield, 3.5% fat-corrected milk yield, and -corrected milk yield. Additionally, PA increased body weight change, but did not affect body condition score change compared with CON. A tendency for a treatment by parity interaction was observed for milk yield due to PA increasing milk yield in multiparous but not in primiparous cows. In addition, we observed interactions between treatment and parity for fat-corrected milk, -corrected milk, and milk fat yield due to PA increasing these variables to a greater extent in multiparous compared with primiparous cows. Interestingly, we observed an interaction between treatment and parity for body weight change, due to PA increasing body weight change in primiparous but not in multiparous cows. The PA treatment increased dry matter and neutral detergent fiber digestibilities compared with CON. Although PA did not affect 18-carbon FA digestibility, compared with CON, PA decreased 16-carbon and total FA digestibilities and increased total FA intake by 470 g/d and absorbed total FA by 316 g/d. We also observed an interaction between treatment and parity for total absorbed FA due to PA increasing it to a greater extent in multiparous than in primiparous cows. Compared with CON, PA increased apparent intake and milk output. We observed an interaction between treatment and parity for milk output due to PA increasing milk output to a greater extent in multiparous than primiparous cows. Additionally, an interaction between treatment and parity was observed for output in body reserves due to PA increasing output in body reserves in primiparous but not in multiparous cows. In conclusion, production responses of dairy cows to PA were consistent throughout the 10-wk treatment period. In addition, PA supplementation interacted with parity, with production responses increased to a greater extent in multiparous than primiparous cows and partitioned to body reserves only increased in primiparous cows.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

A novel physiological role for cardiac myoglobin in lipid metabolism.

Continuous contractile activity of the heart is essential and the required is mostly provided by fatty (FA) oxidation. Myocardial lipid accumulation can lead to pathological responses, however the underlying mechanisms remain elusive. The role of myoglobin in dioxygen binding in cardiomyocytes and oxidative skeletal muscle has widely been appreciated. Our recent work established myoglobin as a protector of cardiac function in hypoxia and disease states. We here unravel a novel role of cardiac myoglobin in governing FA metabolism to ensure the physiological production through β-oxidation, preventing myocardial lipid accumulation and preserving cardiac functions. In vivoH magnetic resonance spectroscopy unveils a 3-fold higher deposition of lipids in mouse hearts lacking myoglobin, which was associated with depressed cardiac function compared to wild-type hearts as assessed by echocardiography. Mass spectrometry reveals a marked increase in tissue triglycerides with preferential incorporation of and oleic acids. Phospholipid levels as well as the metabolome, transcriptome and proteome related to FA metabolism tend to be unaffected by myoglobin ablation. Our results reveal a physiological role of myoglobin in FA metabolism with the lipid accumulation-suppressing effects of myoglobin preventing cardiac lipotoxicity.

Keyword: energy

Two Cockayne Syndrome patients with a novel splice site mutation - clinical and metabolic analyses.

Cockayne Syndrome (CS) is a rare autosomal recessive disorder, which leads to neurodegeneration, growth failure and premature aging. Most of the cases are due to mutations in the ERCC6 gene, which encodes the protein CSB. CSB is involved in several functions including DNA repair and transcription. Here we describe two Danish brothers with CS. Both patients carried a novel splice site mutation (c.2382+2T>G), and a previously described nonsense mutation (c.3259C>T, p.Arg1087X) in a biallelic state. Both patients presented the cardinal features of the disease including microcephaly, congenital cataract and postnatal growth failure. In addition, their fibroblasts were hypersensitive to UV irradiation and exhibited increased superoxide levels in comparison to fibroblasts from healthy age and gender matched individuals. Metabolomic analysis revealed a distinctive metabolic profile in cells from the CS patients compared to control cells. Among others, α-ketoglutarate, hydroxyglutarate and certain amino acids (ornithine, proline and glycine) were reduced in the CS patient fibroblasts, whereas glycolytic intermediates (glucose-6-phosphate and pyruvic ) and fatty acids (, stearic and myristic ) were increased. Our data not only provide additional information to the database of CS mutations, but also point towards targets for potential treatment of this devastating disease.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Potential of oleaginous yeast Trichosporon sp., for conversion of sugarcane bagasse hydrolysate into biodiesel.

This study reports production of microbial oil from a yeast strain Trichosporon sp., (RW) isolated from decayed wood. Preliminary analysis based on fluorescence microscopy and spectroscopy of Nile red stained yeast cells showed accumulation of lipid globules. The potential of the yeast to produce lipids was evaluated on glucose, glycerol and hydrolysate of sugarcane bagasse, where Trichosporon sp. (RW) was found to accumulate 21.45 (59.6%), 18.41 (56%) and 10.25g/l (40.5%) of the lipids after 120h of fermentation at 30°C. FAME analysis of lipids by GC-FID and NMR revealed oleic (18:1) as the major constituent, corresponding to 50.05, 46.48 and 54.66% of the accumulated lipids in glucose, glycerol and hydrolysate grown cultures, respectively. Other accumulated lipids included (16:0), linoleic (18:2) and stearic acids (18:0) in that order. The cetane number of the lipids ranged from 52.39 to 59.57 indicating suitability for biodiesel production.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

[The becoming of reactions of lipolysis in phylogenesis. The and oleic triglycerides as substrates. Insulin, condition of normolipemia and formation of hyper lipoproteinemia type IIb, IV and V].

According to phylogenetic theory of general pathology, when living in ocean all were carnivorous (piscivorous) fatty acids transferring to cells in form of non-polar triglycerides nitially began apoB-48 chylomicrons, continued lipoproteins of very low and low density and fnalized its apoB-100 endocytosis. The fatty acids are transferred by chylomicrons + lipoproteins of very low density + lipoproteins of low density and non-polar triglycerides are hydrolyzed by hepatic glycerolhydrogenase and co-enzyme apoC-III; according WHO classifcation, hyperlipoproteinemia corresponds to type V. On land, in herbivorous who are not yet synthesized insulin, apoB-48 and chylomicrons left process of non-polar triglycerides transferring. In lipoproteins of very low density and lipoproteins of low density, the carnivorous transfer exogenous non-polar triglycerides. The herbivorous also transfer non-polar triglycerides though synthesized by hepatocytes from glucose endogenically. In herbivorous, transferring of non-polar triglycerides prior to synthesis of insulin is forming apoB-100 in composition of lipoproteins of very low density and lipoproteins of low density. The hydrolysis of non-polar triglycerides in lipoproteins of very low density is activated by hepatic glycerol hydrogenase and apoC-III; cells absorb lipoproteins of low density by means of apoB-100 endocytosis. The content on lipoproteins in blood plasma under electrophoresis of lipoproteins corresponds to hepatic glycerol hydrogenase type IIb. In frst and second types of fatty acids transferring in form of triglycerides to lipoproteins of very low density + lipoproteins of low density predominate fatty , triglycerides of the same name and metabolism of fatty acids in vivo. The insulin initiated the third type of transferring of oleic fatty by now to insulin-depended cells only in oleic lipoproteins of very low density; hydrolysis of oleic triglycerides is activated by late in phylogenesis post-heparin hepatic glycerol hydrogenase and apoC-II cofactor. The dynamic apoE is actively bound by apoB-100 forming apoE/B-100 ligand. At later stages of phylogenesis insulin formed fatty acids transferring in form of oleic triglycerides in lipoproteins of very low density of the same name without forming of oleic lipoproteins of low density; the electrophoregram of lipoproteins reflects absence of hepatic glycerol hydrogenase. In phylogenesis three types of fatty acids transferring to triglycerides in composition of lipoproteins formed sequentially: 1) chylomicrons + lipoproteins of very low and density + lipoproteins of low density; 2) lipoproteins of very low density + lipoproteins of low density; 3) only in lipoproteins of very low density. The frst one is specifc to piscivorous (carnivorous) while living in ocean. The second one is implemented by herbivorous while they didn\'t begin to synthesize insulin and hepatocytes not yet transform all endogenous fatty into oleic fatty . Insulin initiated: a) transferring of oleic fatty acids to lipoproteins of very low density without forming oleic lipoproteins of low density; b) highly effective oleic metabolism of fatty acids in vivo: c) becoming of biological function of locomotion. The aphysiological induction by substrate, surplus of fatty acids in food initiate negative alterations in composition of lipoproteins in opposite direction than in case of phylogenesis. When homo sapiens, herbivorous in phylogenesis, begins to misuse carnivorous (meat) food then instead of normolipoproteinemia in blood plasma under electrophoresis of lipoproteins one can initially detect transitory hyperlipoproteinemia type IV and then prolonged hyperlipoproteinemia type IIb. If patient factually passes on to carnivorous diet then hyperlipoproteinemia type V is developing. If content of exogenous fatty in food surpasses physiological capacities of its transferring in oleic triglycerides as palmitoyl-oleyl-palmitate glycerol, triglycerides as oleyl-palmitoyl-palmitate glycerol begin to form and epigenetically aphysiological non-ligand lipoproteins of very low density → lipoproteins of low density are formed. Their circulation in blood is a cause of hypertriglyceridemia, higher level of cholesterol-lipoproteins of low density, compensatory increasing of apoC-III. Then occurs induced by substrate formation of hyperlipoproteinemia initially of type IV, then of type IIb and fnally of type V. The pathogenesis of atherosclerosis and atheromotosis is activated when homo sapiens, herbivorous in phylogenesis, begin to misuse carnivorous food affecting biological functions of trophology, reaction of exotrophy (external nutrition), function of homeostasis, endoecology and function of adaptation. The formation of metabolism if fatty acids instead of oleic one is a cause of chronic defciency of and ATP synthesis in vivo. Insulin activates absorption of glucose by cells with purpose to use it for synthesis of oleic fatty acids. In the frst place, insulin regulates in vivo metabolism of fatty acids and only in second place metabolism of glucose.

Keyword: energy

Kinetics of coffee industrial residue pyrolysis using distributed activation model and components separation of bio-oil by sequencing temperature-raising pyrolysis.

This study was carried out to investigate the kinetics of coffee industrial residue (CIR) pyrolysis, the effect of pyrolysis factors on yield of bio-oil component and components separation of bio-oil. The kinetics of CIR pyrolysis was analyzed using distributed activation model (DAEM), based on the experiments in thermogravimetric analyzer (TGA), and it indicated that the average of activation (E) is 187.86kJ·mol. The bio-oils were prepared from CIR pyrolysis in vacuum tube furnace, and its components were determined by gas chromatography/mass spectrometry (GC-MS). Among pyrolysis factors, pyrolysis temperature is the most influential factor on components yield of bio-oil, directly concerned with the volatilization and yield of components (, linoleic , oleic , octadecanoic and caffeine). Furthermore, a new method (sequencing temperature-raising pyrolysis) was put forward and applied to the components separation of bio-oil. Based on experiments, a solution of components separation of bio-oil was come out.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: energy

Preparation and characterization of 100% bio-based polylactic / microcapsules for thermal storage.

Phase change materials (PCM) have gained extensive attention in thermal storage applications. In this work, microencapsulation of vegetable-derived (PA) in bio-based polylactic (PLA) shell by solvent evaporation and oil-in-water emulsification was investigated. Fourier transform infrared spectroscopy and scanning electron microscopy were conducted to confirm the successful encapsulation of PA in PLA shells. Differential scanning calorimetry was performed to evaluate the thermal properties, thermal reliability, and core content of the fabricated PCM microcapsules (microPCM). Through a series of parametric studies, the effects of PCM and solvent content, oil phase-to-aqueous phase ratio, as well as surfactant type and content on the morphology, particle size, and thermal properties of the PCM microcapsules were investigated. Experimental results showed that PVA was a superior emulsifier to SDS in the emulsion systems being studied. There also existed an optimal PVA concentration to reduce the average size of microPCM. When the PVA concentration was above this optimal level, the emulsifier molecules tend to form micelles among themselves. This led to the adhesion of tiny microspheres on the surface of microPCM as well as larger microPCM. In short, this work has demonstrated the possibility of using the solvent evaporation method to fabricate 100% bio-based PCM-polymer microcapsules for thermal storage applications.

Keyword: energy

Metabolic Profiles Reveal Changes in Wild and Cultivated Soybean Seedling Leaves under Salt Stress.

Clarification of the metabolic mechanisms underlying salt stress responses in plants will allow further optimization of crop breeding and cultivation to obtain high yields in saline-alkali land. Here, we characterized 68 differential metabolites of cultivated soybean (Glycine max) and wild soybean (Glycine soja) under neutral-salt and alkali-salt stresses using gas chromatography-mass spectrometry (GC-MS)-based metabolomics, to reveal the physiological and molecular differences in salt tolerance. According to comparisons of growth parameters under the two kinds of salt stresses, the level of inhibition in wild soybean was lower than in cultivated soybean, especially under alkali-salt stress. Moreover, wild soybean contained significantly higher amounts of phenylalanine, asparagine, citraconic , citramalic , citric and α-ketoglutaric under neutral-salt stress, and higher amounts of , lignoceric , glucose, citric and α-ketoglutaric under alkali-salt stress, than cultivated soybean. Further investigations demonstrated that the ability of wild soybean to salt tolerance was mainly based on the synthesis of organic and amino acids, and the more active tricarboxylic cycle under neutral-salt stress. In addition, the metabolite profiling analysis suggested that the generation from β-oxidation, glycolysis and the citric cycle plays important roles under alkali-salt stress. Our results extend the understanding of mechanisms involved in wild soybean salt tolerance and provide an important reference for increasing yields and developing salt-tolerant soybean cultivars.

Keyword: energy

Dietary sn-2 triacylglycerols reduced faecal lipids, calcium contents and altered lipid metabolism in Sprague-Dawley rats.

In this study, the impact of dietary sn-2 triacylglycerol (sn-2 PTAG) on faecal lipids, calcium excretion and lipid metabolic alternation was investigated in Sprague-Dawley (SD) rats fed with high-fat diet containing either palm olein (PO, sn-2 (PA) of 14.8%), sn-2 PTAG50 (sn-2\u2009PA of 56.4%) or sn-2 PTAG70 (sn-2\u2009PA of 72.4%), respectively. After 4-week feeding period, SD rats fed with sn-2 PTAGs showed reduced faecal soap fatty acids, neutral lipid and calcium excretion compared to those of PO-fed rats, whereas a significant difference was only observed for the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Moreover, dietary sn-2 PTAG70 also showed a significant effect on decreasing serum triacylglycerol (TAG) level, reducing perirenal adipocyte size and regulating lipid metabolism in small intestine and perirenal adipose tissue of SD rats. Significantly increased mRNA levels of genes involved in intestinal lipid anabolism as well as lipid catabolism were both observed in the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Meanwhile, dietary sn-2 PTAG70 also significantly up-regulated lipolysis, mitochondrial fatty oxidation and thermogenesis-related gene and protein levels in perirenal adipose tissue, which might be correlated with the reduced perirenal adipocyte size. Taken together, our findings indicated that sn-2 PTAG70 may have some beneficial effects on intestinal lipid utilisation and lipid metabolic activity for energy supply in visceral adipose tissue.

Keyword: energy

Fatty composition of free-living and parasitic stages of the bovine lungworm Dictyocaulus viviparus.

The development of parasitic nematodes proceeds via multiple stages, often implicating the necessity to adapt to different environments. Especially the transition from free-living to parasitic stages is accompanied by a significant change in the environmental conditions. To shed light on possible adaptations to these transitions, the fatty composition of different developmental stages of the bovine lungworm Dictyocaulus viviparus was investigated. Fatty acids of D. viviparus eggs, the free-living first, second and third larval stage (L1-L3) as well as the parasitic preadult stage and adult male and female worms residing in the lungs of infected hosts were quantified by gas chromatography after transesterification to their fatty methyl esters. The fatty content and diversity were higher in parasitic stages compared to those of free-living larvae. The most prevalent fatty acids in both parasitic and free-living stages were stearic (C18:0), (C16:0), palmitoleic (C16:1) and caprylic (C8:0). A variety of (poly-)unsaturated FAs was found in the parasitic stages and in the eggs, which was similar to the variety of FAs found in bovine surfactant. This finding indicates that parasitic stages of D. viviparus take up FAs from their environment. While eggs contained the highest concentration of fatty acids, a decrease was observed from eggs to L1 and further from L2 to L3. The lowest concentration was found in 38-days-old L3, which suggests that FAs serve as an reserve for the free-living, non-feeding larval stages. The free-living larvae contained mainly saturated fatty acids and only traces of unsaturated fatty acids, which is in contrast to the phospholipid saturation hypothesis of cold tolerance. Instead, a trade-off between desiccation stress and temperature adaptation may favour a higher amount of saturated FAs in the free-living larval stages. Further studies explicitly examining the FA composition of the different classes of lipids are necessary to better describe the adaptative responses of the FA metabolism to different environmental conditions.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

[Ozone oxidizes oleic fatty with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic fatty acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated fatty (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, obesity and nonalcoholic liver disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: energy

Metabolic dependence of cyclosporine\xa0A on cell proliferation of human non‑small cell lung cancer A549 cells and its implication in post‑transplant malignancy.

Cyclosporine\xa0A (CsA), a widely used immunosuppressant to prevent organ transplant rejection, is associated with an increased cancer risk following transplantation, particularly in the lung. However, the underlying mechanisms remain unclear. In the present study, using human non‑small cell lung cancer A549 cells, it was determined that CsA (0.1 or 1\xa0µM) promoted cell proliferation with glucose alone as the source. CsA treatment increased the phosphorylation of protein kinase\xa0B (Akt) and consequently the expression of Cyclin\xa0D1. Inhibiting Akt signaling with the phosphatidylinositol 3‑kinase inhibitor wortmannin prevented this effect. Mechanistically, CsA treatment increased reactive oxygen species (ROS) generation, and the intracellular ROS scavenger N‑acetyl‑cysteine (NAC) attenuated CsA‑induced cell proliferation as well as the activation of Akt/Cyclin\xa0D1 signaling. However, notably, it was demonstrated that CsA treatment decreased cell proliferation and Akt phosphorylation under normal lipid loading. Further investigation indicated that induced excessive generation of ROS, while CsA treatment further stimulated this ROS production. Scavenging intracellular ROS with NAC attenuated the CsA‑mediated inhibition of cell proliferation. Collectively, the results indicated a pleiotropic effect of CsA in the regulation of A549\xa0cell proliferation under different metabolic conditions. This indicated that CsA administration may contribute to increased post‑transplant cancer risk in organ recipients.

Keyword: energy

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers lipid droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in metabolism. However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high-fat diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress lipogenesis. In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic fatty liver disease.

Keyword: energy

Optimization of enzymatic hydrolysis of cellulosic fraction obtained from stranded driftwood feedstocks for lipid production by .

Stranded driftwood feedstocks may represent, after pretreatment with steam explosion and enzymatic hydrolysis, a cheap C-source for producing biochemicals and biofuels using oleaginous yeasts. The hydrolysis was optimized using a response surface methodology (RSM). The solid loading (SL) and the dosage of enzyme cocktail (ED) were variated following a central composite design (CCD) aimed at optimizing the conversion of carbohydrates into lipids (Y) by the yeast DBVPG 5870. A second-order polynomial equation was computed for describing the effect of ED and SL on Y. The best combination (ED\u202f=\u202f3.10%; SL\u202f=\u202f22.07%) for releasing the optimal concentration of carbohydrates which gave the highest predicted Y (27.32%) was then validated by a new hydrolysis. The resulting value of Y (25.26%) was close to the theoretical maximum value. Interestingly, fatty profile achieved under the optimized conditions was similar to that reported for palm oil.

Keyword: energy

Functional lipidomics: impairs hepatocellular carcinoma development by modulating membrane fluidity and glucose metabolism.

Lipids are essential cellular components and sources of living organisms, and altered lipid composition is increasingly recognized as a signature of cancer. We performed lipidomic analysis in a series of hepatocellular carcinoma (HCC) cells and identified over 1,700 intact lipids originating from three major lipid categories. Comparative lipidomic screening revealed that 93 significantly changed lipids and decreased acyl (C16:0)-containing glycerophospholipids were positively associated with metastatic abilities of HCC cells. Furthermore, both in vitro and in vivo experiments demonstrated that C16:0 incubation specifically reduced malignant cell proliferation, impaired cell invasiveness, and suppressed tumor growth in mouse xenograft models. Biochemical experiments demonstrated that C16:0 treatment decreased cell membrane fluidity and limited glucose metabolism. A phosphoproteomics approach further revealed such C16:0 incubation attenuated phosphorylation levels of mammalian target of rapamycin (mTOR) and signal transducer and activator of transcription 3 (STAT3) pathway proteins. Multiple reaction monitoring analysis of 443 lipid molecules showed 8 reduced C16:0-containing lipids out of total 10 altered lipids when cancer tissues were compared with adjacent nontumor tissues in a cohort of clinical HCC specimens (P < 0.05).These data collectively demonstrate the biomedical potential of using altered lipid metabolism as a diagnostic marker for cancerous cells and open an opportunity for treating aggressive HCCs by targeting altered C16:0 metabolism. (Hepatology 2017;66:432-448).© 2017 by the American Association for the Study of Liver Diseases.

Keyword: energy

Single Cell Oil Production from Undetoxified L. hydrolysate by .

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, . is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast from undetoxified lignocellulosic hydrolysates of steam-pretreated . The growth of was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty profile of triglycerides contained 27% , 33% oleic and 32% linoleic . These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Keyword: energy

Good Fats versus Bad Fats: A Comparison of Fatty Acids in the Promotion of Insulin Resistance, Inflammation, and Obesity.

Recently, debate has erupted in both the scientific community and throughout the lay public around whether a low-fat or low-carbohydrate diet is better for weight loss. In other words, is it better to cut fat or cut carbohydrate for weight loss. However, going beyond this debate (fat versus carbohydrate), are questions around whether certain fatty acids are worse for promoting insulin resistance, inflammation, and obesity. The overall evidence in the literature suggests that medium-chain saturated fats (such as lauric , found in coconut oil) and monounsaturated fat (oleic , found in olive oil) are less likely to promote insulin resistance, inflammation, and fat storage compared to long-chain saturated fatty acids (such as stearic found in large quantities in butter, but particularly found in palm oil) especially when consumed on top of a diet moderate in refined carbohydrates. Compared to long-chain saturated fats, lauric and oleic have an increased fatty oxidation rate, are more likely to be burned for and less likely to be stored in adipose tissue, and thus promote increased expenditure. Omega-6 polyunsaturated fatty acids (PUFAs), such as linoleic , as found in vegetable oils may contribute to obesity, whereas omega-3 PUFA may be protective. Importantly, both olive oil as part of a Mediterranean diet, and omega-3 from fish and fish oil have been proven to reduce risk of cardiovascular (CV) events.

Keyword: energy

Chemical and Nutritional Composition of Terminalia ferdinandiana (Kakadu Plum) Kernels: A Novel Nutrition Source.

(Kakadu plum) is a native Australian fruit. Industrial processing of fruits into puree generates seeds as a by-product, which are generally discarded. The aim of our present study was to process the seed to separate the kernel and determine its nutritional composition. The proximate, mineral and fatty compositions were analysed in this study. Kernels are composed of 35% fat, while proteins account for 32% dry weight (DW). The content and fiber were 2065 KJ/100 g and 21.2% DW, respectively. Furthermore, the study showed that kernels were a very rich source of minerals and trace elements, such as potassium (6693 mg/kg), calcium (5385 mg/kg), iron (61 mg/kg) and zinc (60 mg/kg) DW, and had low levels of heavy metals. The fatty composition of the kernels consisted of omega-6 fatty , linoleic (50.2%), monounsaturated oleic (29.3%) and two saturated fatty acids namely (12.0%) and stearic (7.2%). The results indicate that kernels have the potential to be utilized as a novel protein source for dietary purposes and non-conventional supply of linoleic, and oleic acids.

Keyword: energy

Fabrication and Performance of Composite Microencapsulated Phase Change Materials with Ethyl Ester as Core.

Microencapsulation of phase change materials (PCMs) could prevent the leakage of PCMs during solid⁻liquid phase change process. However, their applications are mainly limited by the compactness and thermal stability of the traditional polyurea shell microcapsules. To increase the thermal compactness and thermal stability of PCM microcapsules, tetraethylorthosilicate (TEOS) was employed to form polymer/SiO₂ composite shells to enhance the mechanical performance of polyurea and polyurethane microcapsule via interfacial polymerization and in situ polymerization. The morphology and chemical components of the microcapsules were characterized by field-emission scanning electron microscope (FE-SEM) and Fourier transform infrared (FT-IR) spectroscopy, respectively. The thermal properties of the microcapsules were investigated by differential scanning calorimetry (DSC) and thermal gravity analysis (TGA). The results showed the smoothness and compactness of both polyurea⁻SiO₂ and polyurethane⁻SiO₂ microcapsules enhanced slightly, when compared with that without TEOS addition. Moreover, the SiO₂ composite shell had good effect on thermal compactness, as the weight loss rate of polyurea⁻SiO₂ microcapsules and polyurethane⁻SiO₂ microcapsules decreased 3.5% and 4.1%, respectively.

Keyword: energy

Green preparation of seaweed-based silver nano-liquid for cotton pathogenic fungi management.

Silver nanoparticles (Ag NPs) were synthesised using the crude ethyl acetate extracts of and evaluated their bioefficacy against two crop-damaging pathogens. The sets of lattice planes in the XRD spectrum for the Ag NPs were indexed to the 111, 200, 220 and 311 orientations and support the crystalline nature of the Ag NPs. The 3414 and 2968\u2005cm peaks were observed in crude algal thallus extract and they were characteristic of terpenoids. Further, a peak at 1389\u2005cm was observed as fatty acids. The marine macroalgae terpenoids and acted as reducing agent and stabiliser, respectively. The size (3 and 50\u2005nm) and shape (spherical) of Ag NPs were recorded. The -dispersive X-ray spectroscopy analysis exemplified the presence of silver in its elemental nature. Moreover, Ag NPs were effective against two cotton phytopathogens namely f.sp. (FOV) and pv. malvacearum (XAM). The minimum inhibitory concentration was found to be 80.0 and 43.33\u2005μg ml against FOV and XAM, respectively. Results confirmed the anti-microbial activity of green nanoparticles against select pathogens and suggest their possible usage in developing antifungal agents for controlling destructive pathogens in a cotton agroecosystem.

Keyword: energy

Dehydration Causes Increased Reliance on Protein Oxidation in Mice: A Test of the Protein-for-Water Hypothesis in a Mammal.

During fasting, animals rely on a mixture of fats, carbohydrates, and proteins that are derived solely from endogenous sources. The relative contributions of these metabolic fuels chiefly depend on the duration of the fast, but other factors including previous diet, environmental temperature, and activity level can modulate the fuel mixture. It has long been held that endogenous proteins are spared from catabolism until the final stages of prolonged fasting and contribute a significant proportion of once the other metabolic fuels have been depleted. However, evidence is mounting that protein is catabolized supplemental to fat metabolism under some circumstances. This has been shown in migratory birds that exhibit dramatic reductions in lean mass during flights. One hypothesis to explain this seemingly maladaptive metabolic strategy is that the catabolism and oxidation of protein in situ yields five times more metabolic water than that generated through fat oxidation alone. Recent support for this hypothesis is that birds benefit from such a strategy due to their uricotelic nature. However, it remains unclear whether ureotelic mammals would also employ this strategy. Here we test the protein-for-water hypothesis in resting laboratory mice subjected to water deprivation during fasting while we tracked rates of protein and lipid catabolism using endogenously incorporated C-leucine and C-. We found no differences in instantaneous leucine oxidation; however, cumulative differences in instantaneous leucine oxidation ultimately resulted in a higher total leucine oxidation after 72 h of fasting in water-deprived animals. We also found that lipid oxidation was 8% higher in the hydrated mice, but the difference was not significant presumably because of a concomitant reduction in metabolic rates of the water-deprived mice. Our results indicate that mammals do increase rates of protein catabolism during dehydration but to a lesser degree than birds. The ability of mammals to produce highly concentrated urine and their lower inherent rates of protein turnover apparently preclude mammals from taking full advantage of the protein-for-water strategy during fasting under dehydrating conditions.

Keyword: energy

Fatty profiles of the European migratory common noctule bat (Nyctalus noctula).

In animals, fatty acids (FA) are essential as structural components in membranes and for storage in adipocytes. Here, we studied the relative proportions of FA in a mammal with extreme changes in metabolic rates. Common noctule bats (Nyctalus noctula) switch from energetically demanding long-distance migration at high metabolic rates to regular torpor with extremely low metabolic rates. We found that composition of FA categories differed between adipose tissue types (white adipose tissue (WAT) vs brown adipose tissue (BAT)) and muscle tissue types (skeletal vs heart), but not between sexes. We found oleic to be the most abundant FA in all studied tissues. Concentrations of polyunsaturated FA (PUFA) were not always higher in muscular tissue compared with adipocyte tissue, even though high concentrations of PUFA are considered beneficial for low body temperatures in torpor. In all tissues, we observed a high content in monounsaturated fatty acids (MUFA), possibly to compensate for a low PUFA content in the diet. Ratios of ω6/ω3 were lower in the heart than in skeletal muscles of common noctules. Three FA (, oleic, and linoleic ) accounted for about 70% of the FA in adipose tissue, which is similar to proportions observed in migrating birds, yet migrating birds generally have a higher PUFA content in muscle and adipose tissues than bats. Bats seem to contrast with other mammals in having a high MUFA content in all tissues. We conclude that FA profiles of bats differ largely from those of most cursorial mammals and instead are-with the exception of MUFA-similar to those of migrating birds.

Keyword: energy

Altering the ratio of dietary C16:0 and cis-9 C18:1 interacts with production level in dairy cows: Effects on production responses and partitioning.

The objective of our study was to evaluate the effects of altering the dietary ratio of (C16:0) and oleic (cis-9 C18:1) acids on nutrient digestibility, partitioning, and production responses of lactating dairy cows. Cows were blocked by milk yield and assigned to 3 groups (12 cows per group) in a main plot: low (45.2 ± 1.7 kg/d), medium (53.0 ± 1.6 kg/d), and high (60.0 ± 1.9 kg/d). Within each production group, a truncated Latin square arrangement of fatty (FA) treatments was used in 2 consecutive 35-d periods. The FA treatments supplemented at 1.5% of diet dry matter were (1) 80:10 (80% C16:0 + 10% cis-9 C18:1), (2) 73:17 (73% C16:0 + 17% cis-9 C18:1), (3) 66:24 (66% C16:0 + 24% cis-9 C18:1), and (4) 60:30 (60% C16:0 + 30% cis-9 C18:1). Treatment × production group interactions were observed for yields of milk, fat-corrected milk, -corrected milk, milk fat, milk protein, and milk lactose and partitioned to milk. Increasing cis-9 C18:1 in FA treatments reduced fat-corrected milk, -corrected milk, and milk output in low-producing cows but increased these in high-producing cows. Increasing cis-9 C18:1 in FA treatments did not affect milk yield, milk protein yield, and milk lactose yield in low- and medium-producing cows but increased these in high-producing cows. Regardless of production level, there was no effect of treatments on dry matter intake; however, increasing cis-9 C18:1 in FA treatments increased body weight change and body condition score change. Increasing cis-9 C18:1 in FA treatments increased total FA digestibility due to a linear increase in 16- and 18-carbon FA digestibilities. Interactions between FA treatments and production level were observed for the yield of milk fat and milk FA sources. In low-producing cows, increasing cis-9 C18:1 in FA treatments decreased milk fat yield due to a decrease in de novo and mixed milk FA without changes in preformed milk FA. In contrast, in high-producing cows, increasing cis-9 C18:1 in FA treatments increased milk fat yield due to an increase in de novo and preformed milk FA. Our results indicate that high-producing dairy cows (averaging 60 kg/d) responded better to a fat supplement containing more cis-9 C18:1, whereas low-producing cows (averaging 45 kg/d) responded better to a supplement containing more C16:0.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Linoleic rescues microglia inflammation triggered by saturated fatty .

Elevated saturated free fatty levels during over-nutrition lead to hypothalamic inflammation, which perturbs homeostasis. Whether brain-derived metabolites are coupled to the development of obesity pathogenesis during the early over-nutrition period has not been thoroughly investigated. In this study, we found increased linoleic , an unsaturated fatty , in both the whole brain and hypothalamus of mice fed a high-fat diet for 4 weeks. Furthermore, we observed that linoleic effectively reversed the inflammatory responses induced by treatment in microglial cells. Collectively, this study suggests the reversible function of linoleic on brain inflammation in association with microglial activation during short-term exposure to a high-fat diet.Copyright © 2019. Published by Elsevier Inc.

Keyword: energy

Human embryonic stem cell-derived cardiomyocytes as an in vitro model to study cardiac insulin resistance.

Patients with type 2 diabetes (T2D) and/or insulin resistance (IR) have an increased risk for the development of heart failure (HF). Evidence indicates that this increased risk is linked to an altered cardiac substrate preference of the insulin resistant heart, which shifts from a balanced utilization of glucose and long-chain fatty acids (FAs) towards an almost complete reliance on FAs as main fuel source. This shift leads to a loss of endosomal proton pump activity and increased cardiac fat accumulation, which eventually triggers cardiac dysfunction. In this review, we describe the advantages and disadvantages of currently used in vitro models to study the underlying mechanism of IR-induced HF and provide insight into a human in vitro model: human embryonic stem cell-derived cardiomyocytes (hESC-CMs). Using functional metabolic assays we demonstrate that, similar to rodent studies, hESC-CMs subjected to 16h of high palmitate (HP) treatment develop the main features of IR, i.e., decreased insulin-stimulated glucose and FA uptake, as well as loss of endosomal acidification and insulin signaling. Taken together, these data propose that HP-treated hESC-CMs are a promising in vitro model of lipid overload-induced IR for further research into the underlying mechanism of cardiac IR and for identifying new pharmacological agents and therapeutic strategies. This article is part of a Special issue entitled Cardiac adaptations to obesity, diabetes and insulin resistance, edited by Professors Jan F.C. Glatz, Jason R.B. Dyck and Christine Des Rosiers.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

Adaptations of hepatic lipid metabolism and mitochondria in dairy cows with mild fatty liver.

The inevitable deficiency in nutrients and at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. Fatty liver is one of the main health disorders after parturition. Therefore, to investigate changes in hepatic lipid metabolic status and mitochondria in dairy cows with mild fatty liver, liver and blood samples were collected from healthy cows (n = 15) and cows with mild fatty liver (n = 15). To determine the effects of acids (PA), one of the major component of fatty acids, on lipid metabolism and mitochondria in vitro, calf hepatocytes were isolated from healthy calves and treated with various concentrations of PA (0, 50, 100, and 200 μM). Dairy cows with mild fatty liver displayed hepatic lipid accumulation. The protein levels of sterol regulatory element-binding protein 1c (SREBP-1c) and peroxisome proliferator-activated receptor-α (PPARα) and mRNA levels of acetyl CoA carboxylase 1 (ACC1), fatty synthase (FAS), acyl-CoA oxidase (ACO), and carnitine palmitoyltransferase 1A (CPT1A) were significantly higher in dairy cows with mild fatty liver than in control cows. The hepatic mitochondrial DNA content, mRNA levels of oxidative phosphorylation complexes I to V (CO 1-V), protein levels of cytochrome c oxidase subunit IV (COX IV), voltage dependent anion channel 1 (VDAC1), peroxisome proliferator activated receptor-γ coactivator-1α (PGC-1α) and nuclear respiratory factor 1 (NRF1), and adenosine triphosphate (ATP) content were all markedly increased in the liver of dairy cows with mild fatty liver compared with healthy cows. The PA treatment significantly increased lipid accumulation; protein levels of SREBP-1c and PPARα; and mRNA levels of ACC1, FAS, ACO, and CPT1A in calf hepatocytes. Moreover, the mitochondrial DNA content, mRNA levels of CO 1-V, protein levels of COX IV, VDAC1, PGC-1α, NRF1, mitochondrial transcription factor A, and ATP content were significantly increased in PA-treated hepatocytes compared with control hepatocytes. The protein level of mitofusin-2 was significantly decreased in PA-treated groups. In conclusion, lipid synthesis and oxidation, number of mitochondria, and ATP production were increased in the liver of dairy cows with mild fatty liver and PA-treated calf hepatocytes. These changes in hepatic mitochondria and lipid metabolism may be the adaptive mechanism of dairy cows with mild fatty liver.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Exploring the potency of integrating semi-batch operation into lipid yield performance of Chlamydomonas sp. Tai-03.

Third generation biofuels, also known as microalgal biofuels, are promising alternatives to fossil fuels. One attractive option is microalgal biodiesel as a replacement for diesel fuel. Chlamydomonas sp. Tai-03 was previously optimized for maximal lipid production for biodiesel generation, achieving biomass growth and productivity of 3.48\u202f±\u202f0.04\u202fg/L and 0.43\u202f±\u202f0.01\u202fg/L/d, with lipid content and productivity of 28.6\u202f±\u202f1.41% and 124.1\u202f±\u202f7.57\u202fmg/L/d. In this study, further optimization using 5% CO concentration and semi-batch operation with 25% medium replacement ratio, enhanced the biomass growth and productivity to 4.15\u202f±\u202f0.12\u202fg/L and 1.23\u202f±\u202f0.02\u202fg/L/d, with lipid content and productivity of 19.4\u202f±\u202f2.0% and 239.6\u202f±\u202f24.8\u202fmg/L/d. The major fatty methyl esters (FAMEs) were (C16:0), oleic (C18:1), and linoleic (C18:2). These short-chain FAMEs combined with high growth make Chlamydomonas sp. Tai-03 a suitable candidate for biodiesel synthesis.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: energy

Short communication: Comparison of a -enriched triglyceride supplement and calcium salts of palm fatty acids supplement on production responses of dairy cows.

The objective of our study was to evaluate the effects of feeding a -enriched triglyceride supplement or a calcium salts of palm fatty (FA) supplement on nutrient digestibility and production responses of mid-lactation dairy cows. Fifteen Holstein cows (139 ± 39 d in milk) were randomly assigned to treatment sequence in a 3 × 3 Latin square design. Treatments were a control diet (CON; no fat supplement) and 1.5% of FA added either as a -enriched triglyceride supplement (PA-TG) or as calcium salts of palm FA supplement (Ca-FA). Fat-supplemented treatments did not affect dry matter intake (DMI) compared with CON, but Ca-FA reduced DMI compared with PA-TG. Compared with CON, fat-supplemented treatments increased 18-carbon FA digestibility by 2.0 percentage units but did not affect digestibility of total FA or 16-carbon FA. Compared with Ca-FA, PA-TG reduced total FA digestibility by 8.7 percentage units due to a decrease in 16-carbon FA digestibility (21.7 percentage units). Both fat supplements increased neutral detergent fiber (NDF) digestibility compared with CON (3.90 percentage units), and PA-TG tended to increase NDF digestibility by 1.60 percentage units compared with Ca-FA. Compared with CON, fat-supplemented treatments increased milk yield (1.05 kg/d), 3.5% fat-corrected milk yield (2.20 kg/d), and -corrected milk yield (1.80 kg/d). Also, PA-TG increased milk fat yield (50 g/d) and milk output (1.0 Mcal/d) and tended to increase milk fat content (0.07 percentage units) and -corrected milk yield (1.0 kg/d) compared with Ca-FA. Fat-supplemented treatments reduced the yield of de novo milk FA (23 g/d) and increased the yields of mixed (43 g/d) and preformed (52 g/d) milk FA compared with CON. The PA-TG treatment increased the yield of 16-carbon (66 g/d) milk FA compared with Ca-FA, whereas Ca-FA increased the yield of preformed (60 g/d) milk FA. Fat-supplemented treatments increased intake of net for lactation by 1.80 Mcal/d, milk output by 1.30 Mcal/d, and in body reserves by 0.30 Mcal/d compared with CON. The Ca-FA treatment increased allocated to body reserves (0.60 Mcal/d), partitioning toward body reserves (1.20 percentage units), and body condition score change (0.06 units), and tended to increase body weight change (0.16 kg/d) and body condition score (0.08 units) compared with PA-TG. In conclusion, feeding a -enriched triglyceride supplement increased milk output due to increased yields of milk and milk fat, whereas feeding a calcium salts of palm FA supplement increased FA digestibility and partitioned to body reserves.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the metabolism of fatty acids (FA) and only in the second turn in glucose metabolism; regulation of FA metabolism in cells started millions of years earlier than that of glucose metabolism. Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral fat cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA metabolism instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, metabolic syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: energy

Resveratrol Ameliorates Lipid Droplet Accumulation in Liver Through a SIRT1/ ATF6-Dependent Mechanism.

Lipid droplets (LDs) are dynamic organelles that store neutral lipids during times of excess, and an increased accumulation of LDs in the liver is closely linked to hepatic steatosis. Our previous studies suggested that resveratrol (RSV) supplement could improve hepatic steatosis, but the underlying mechanism, particularly which related to LD accumulation, has not yet been elucidated.A high-fat diet (HFD) and were used to induce hepatic steatosis in mouse liver and hepatocytes, respectively. The effects of RSV on LD accumulation were analyzed in vivo and in vitro. The effects of RSV on the expression levels of LD-associated genes (ATF6, Fsp27β/CIDEC, CREBH, and PLIN1) were measured by qRT-PCR and western blot assays, followed by KD or overexpression of SIRT1 and ATF6 with small interfering RNAs or overexpressed plasmids, respectively. The dual luciferase reporter assay, chromatin immunoprecipitation assay, coimmunoprecipitation, and proximity ligation assay were utilized to clarify the mechanism of transcriptional regulation and possible interaction between SIRT1 and ATF6.There was a significant increase in the accumulation of LDs in liver and hepatocytes during the process of HFD-induced steatosis, respectively, which was significantly inhibited by RSV supplementation. RSV notably activated SIRT1 expression and decreased the expression levels of ATF6, Fsp27β/CIDEC, CREBH, and PLIN1, which are associated with LD accumulation. Interestingly, the inhibitory effects of RSV on LD accumulation and the associated expression of genes in hepatocytes were abrogated or strengthened with SIRT1 silencing or overexpression, respectively. On the contrary, the benefits of RSV in hepatocytes were eliminated or aggravated when transfected with the overexpressed ATF6 or ATF6 siRNA, respectively. Furthermore, we found that RSV stimulated SIRT1 expression significantly, which was followed by increased deacetylation and inactivation of ATF6, resulting in a positive feedback loop for SIRT1 transcription associated with ATF6 binding to the SIRT1 promoter region.Taken together, these findings indicate that RSV supplementation improves hepatic steatosis by ameliorating the accumulation of LDs, and this might be partially mediated by a SIRT1/ATF6-dependent mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: energy

Sodium-carboxylate contact ion pair formation induces stabilization of monolayers at high pH.

Sea spray aerosols (SSA) are known to have an organic coating that is mainly composed of fatty acids. In this study, the effect of pH and salt on the stability and organization of a (PA) monolayer is investigated by surface vibrational spectroscopy and molecular dynamics simulations. Results indicate that alkyl chain packing becomes more disordered as the carboxylic headgroup becomes deprotonated. This is associated with packing mismatch of charged and neutral species as charged headgroups penetrate deeper into the solution phase. At pH 10.7, when the monolayer is ∼99% deprotonated, palmitate (PA) molecules desorb and solubilize into the bulk solution where there is spectroscopic evidence for aggregate formation. Yet, addition of 100 mM NaCl to the bulk solution is found to drive PA molecules to the aqueous surface. Free calculations show that PA molecules become stabilized within the interface with increasing NaCl concentration. Formation of contact -COO:Na pairs alters the hydration state of PA headgroups, thus increasing the surface propensity. As salts are highly concentrated in SSA, these results suggest that deprotonated fatty acids may be found at the air-aqueous interface of aerosol particles due to sea salt\'s role in surface stabilization.

Keyword: energy

The selective production of jet fuel range alkanes via the catalytic upgrading of over Co/HMCM-49 catalysts.

The complete deoxygenation of followed by simultaneous hydro-isomerization and appropriate hydrocracking were achieved over Co/HMCM-49 bi-functional catalysts with the special structure of 12 MR cups and a low Si/Al ratio. This offered new insights into the design of bi-functional catalysts for carbon rearrangements during the upgrading of to jet fuel range alkanes.

Keyword: energy

Own-Synthetize Nanoparticles to Develop Nano-Enhanced Phase Change Materials (NEPCM) to Improve the Efficiency in Buildings.

The use of adequate thermal storage (TES) systems is an opportunity to increase efficiency in the building sector, and so decrease both commercial and residential consumptions. Nano-enhanced phase change materials (NEPCM) have attracted attention to address one of the crucial barriers (i.e. low thermal conductivity) to the adoption of phase change materials (PCM) in this sector. In the present study two PCM based on fatty acids, capric and , were nano-enhanced with low contents (1.0 wt.%, 1.5 wt.% and 3.0 wt.%) of copper (II) oxide (CuO) nanoparticles. Copper (II) oxide (CuO) was synthesized via coprecipitation method obtaining 60⁻120 nm diameter sized nanoparticles. Thermal stability and high thermal conductivity were observed for the nano-enhanced phase change materials (NEPCM) obtained. Experimental results revealed remarkable increments in NEPCM thermal conductivity, for instance thermal conductivity was increased up to 60% with the addition of 3 wt.% CuO nanoparticles. Moreover, CuO nanoparticles sedimentation velocity decreases when increasing its content.

Keyword: energy

stimulates metabolism and inhibits insulin/PI3K/AKT signaling in differentiated human neuroblastoma cells: The role of mTOR activation and mitochondrial ROS production.

The high consumption of saturated lipids has been largely associated with the increasing prevalence of metabolic diseases. In particular, saturated fatty acids such as (PA) have been implicated in the development of insulin resistance in peripheral tissues. However, how neurons develop insulin resistance in response to lipid overload is not fully understood. Here, we used cultured rat cortical neurons and differentiated human neuroblastoma cells to demonstrate that PA blocks insulin-induced metabolic activation, inhibits the activation of the insulin/PI3K/Akt pathway and activates mTOR kinase downstream of Akt. Despite the fact that fatty acids are not normally used as a significant source of fuel by neural cells, we also found that short-term neuronal exposure to PA reduces the NAD/NADH ratio, indicating that PA modifies the neuronal balance. Finally, inhibiting mitochondrial ROS production with mitoTEMPO prevented the deleterious effect of PA on insulin signaling. This work provides novel evidence of the mechanisms behind saturated fatty -induced insulin resistance and its metabolic consequences on neuronal cells.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Seed traits, fatty profile and genetic diversity assessment in Pongamia pinnata (L.) Pierre germplasm.

Phenotypic variation of important seed traits like seed length, seed breadth, seed thickness, 100 seed weight and seed oil content were recorded in a total of 157 collected accessions of Pongamia. Out of these, fatty profiles of 38 accessions selected based on their high and low oil content was analyzed. Fatty profile revealed high variability in stearic, oleic and linoleic which varied from 0.42 to 10.61\xa0%, 34.34 to 74.58\xa0%, and 7.00 to 31.28\xa0% respectively. Variations in and linolenic were small. Iodine value, saponification number and cetane number (CN) of fatty methyl esters (FAME) of seed oil ranges from 186.99 to 201.25, 81.13 to 108.19 and 46.16 to 56.47 respectively. Fatty compositions, degree of unsaturation and CN are the important parameters, which are used to determine quality of FAME were used as biodiesel. Some of the Pongamia accessions identified were higher in oil content while some accessions showed higher degree of unsaturation and a few of them had CN values higher than 55. Genetic diversity analysis with six TE-AFLP primers generated a total of 334 bands out of which 174 (52.10\xa0%) were polymorphic. The genetic similarity ranged from 0.11 to 0.47. These findings clearly showed high level of genetic diversity and all economically desirable traits were not present in a single genotype of Pongamia. All these traits could be selected from these CPTs and transfer to a single elite variety through selection and breeding programme and could be utilized for large scale multiplication and plantation to produce high quantity and quality biodiesel in future.

Keyword: energy

Wax Ester Fermentation and Its Application for Biofuel Production.

In Euglena cells under anaerobic conditions, paramylon, the storage polysaccharide, is promptly degraded and converted to wax esters. The wax esters synthesized are composed of saturated fatty acids and alcohols with chain lengths of 10-18, and the major constituents are myristic and myristyl alcohol. Since the anaerobic cells gain ATP through the conversion of paramylon to wax esters, the phenomenon is named "wax ester fermentation". The wax ester fermentation is quite unique in that the end products, i.e. wax esters, have relatively high molecular weights, are insoluble in water, and accumulate in the cells, in contrast to the common fermentation end products such as lactic and ethanol.A unique metabolic pathway involved in the wax ester fermentation is the mitochondrial fatty synthetic system. In this system, fatty are synthesized by the reversal of β-oxidation with an exception that trans-2-enoyl-CoA reductase functions instead of acyl-CoA dehydrogenase. Therefore, acetyl-CoA is directly used as a C donor in this fatty synthesis, and the conversion of acetyl-CoA to malonyl-CoA, which requires ATP, is not necessary. Consequently, the mitochondrial fatty synthetic system makes possible the net gain of ATP through the synthesis of wax esters from paramylon. In addition, acetyl-CoA is provided in the anaerobic cells from pyruvate by the action of a unique enzyme, oxygen sensitive pyruvate:NADP oxidoreductase, instead of the common pyruvate dehydrogenase multienzyme complex.Wax esters produced by anaerobic Euglena are promising biofuels because myristic (C) in contrast to other algal produced fatty acids, such as (C) and stearic (C), has a low freezing point making it suitable as a drop-in jet fuel. To improve wax ester production, the molecular mechanisms by which wax ester fermentation is regulated in response to aerobic and anaerobic conditions have been gradually elucidated by identifying individual genes related to the wax ester fermentation metabolic pathway and by comprehensive gene/protein expression analysis. In addition, expression of the cyanobacterial Calvin cycle fructose-1,6-bisphosphatase/sedohepturose-1,7-bisphosphatase, in Euglena provided photosynthesis resulting in increased paramylon accumulation enhancing wax ester production. This chapter will discuss the biochemistry of the wax ester fermentation, recent advances in our understanding of the regulation of the wax ester fermentation and genetic engineering approaches to increase production of wax esters for biofuels.

Keyword: energy

Bi-allelic mutations of LONP1 encoding the mitochondrial LonP1 protease cause pyruvate dehydrogenase deficiency and profound neurodegeneration with progressive cerebellar atrophy.

LonP1 is crucial for maintaining mitochondrial proteostasis and mitigating cell stress. We identified a novel homozygous missense LONP1 variant, c.2282 C\xa0>\xa0T, (p.Pro761Leu), by whole-exome and Sanger sequencing in two siblings born to healthy consanguineous parents. Both siblings presented with stepwise regression during infancy, profound hypotonia and muscle weakness, severe intellectual disability and progressive cerebellar atrophy on brain imaging. Muscle biopsy revealed the absence of ragged-red fibers, however, scattered cytochrome c oxidase-negative staining and electron dense mitochondrial inclusions were observed. Primary cultured fibroblasts from the siblings showed normal levels of mtDNA and mitochondrial transcripts, and normal activities of oxidative phosphorylation complexes I through V. Interestingly, fibroblasts of both siblings showed glucose-repressed oxygen consumption compared to their mother, whereas galactose and utilization were similar. Notably, the siblings\' fibroblasts had reduced pyruvate dehydrogenase (PDH) activity and elevated intracellular lactate:pyruvate ratios, whereas plasma ratios were normal. We demonstrated that in the siblings\' fibroblasts, PDH dysfunction was caused by increased levels of the phosphorylated E1α subunit of PDH, which inhibits enzyme activity. Blocking E1α phosphorylation activated PDH and reduced intracellular lactate concentrations. In addition, overexpressing wild-type LonP1 in the siblings\' fibroblasts down-regulated phosphoE1α. Furthermore, in vitro studies demonstrated that purified LonP1-P761L failed to degrade phosphorylated E1α, in contrast to wild-type LonP1. We propose a novel mechanism whereby homozygous expression of the LonP1-P761L variant leads to PDH deficiency and metabolism dysfunction, which promotes severe neurologic impairment and neurodegeneration.

Keyword: energy

Flaxseed oil ameliorated high-fat-diet-induced bone loss in rats by promoting osteoblastic function in rat primary osteoblasts.

α-Linolenic (ALA) is a plant-derived omega-3 unsaturated fatty that is rich in flaxseed oil (FO). The effect of FO on bone health is controversial. This study aims to evaluate the effect of FO on bone damage induced by a high-fat diet (HFD) and to explore the possible mechanism.Male Sprague-Dawley rats were fed a normal control diet (NC, 10% fat), FO diet (NY, 10% fat), HFD (60% fat), or HFD containing 10% FO (HY, 60% fat) for 22\u2009weeks. Micro CT and three-point bending tests were conducted to evaluate bone microstructure and biomechanics. Serum was collected for the detection of ALP, P1NP, and CTX-1. Rat primary osteoblasts (OBs) were treated with different concentrations of ALA with or without (PA) treatment. The ALP activity, osteogenic-related gene and protein expression were measured.Rats in the HFD group displayed decreased biomechanical properties, such as maximum load, maximum fracture load, ultimate tensile strength, stiffness, absorption, and elastic modulus, compared with the NC group ( \xa0<\u20090.05). However, HY attenuated the HFD-induced decreases in bone biomechanical properties, including maximum load, maximum fracture load, and ultimate tensile strength (\u2009<\u20090.05). Trabecular bone markers such as trabecular volume bone mineral density (Tb. vBMD trabecular bone volume/total volume (Tb. BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) were decreased, trabecular separation (Tb. Sp) and the structure model index (SMI) were increased in the HFD group compared with the NC group, and all parameters were remarkably improved in the HY group compared to the HFD group (p\u2009<\u20090.05). However, cortical bone markers such as cortical volume bone mineral density (Ct. vBMD), cortical bone volume/total volume (Ct. BV/TV) and cortical bone thickness (Ct. Th) were not significantly different among all groups. Moreover, the serum bone formation markers ALP and P1NP were higher and the bone resorption marker CTX-1 was lower in the HY group compared with levels in the HFD group. Compared with the NC group, the NY group had no difference in the above indicators. In rat primary OBs, PA treatment significantly decreased ALP activity and osteogenic gene and protein (β-catenin, RUNX2, and osterix) expression, and ALA dose-dependently restored the inhibition induced by PA.FO might be a potential therapeutic agent for HFD-induced bone loss, most likely by promoting osteogenesis.© The Author(s). 2019.

Keyword: energy

Role of the saturated fatty palmitate in the interconnected hypothalamic control of homeostasis and biological rhythms.

The brain, specifically the hypothalamus, controls whole body and glucose homeostasis through neurons that synthesize specific neuropeptides, whereas hypothalamic dysfunction is linked directly to insulin resistance, obesity, and type 2 diabetes mellitus. Nutrient excess, through overconsumption of a Western or high-fat diet, exposes the hypothalamus to high levels of free fatty acids, which induces neuroinflammation, endoplasmic reticulum stress, and dysregulation of neuropeptide synthesis. Furthermore, exposure to a high-fat diet also disrupts normal circadian rhythms, and conversely, clock gene knockout models have symptoms of metabolic disorders. While whole brain/animal studies have provided phenotypic end points and important clues to the genes involved, there are still major gaps in our understanding of the intracellular pathways and neuron-specific components that ultimately control circadian rhythms and homeostasis. Because of its complexity and heterogeneous nature, containing a diverse mix cell types, it is difficult to dissect the critical hypothalamic components involved in these processes. Of significance, we have the capacity to study these individual components using an extensive collection of both embryonic- and adult-derived, immortalized hypothalamic neuronal cell lines from rodents. These defined neuronal cell lines have been used to examine the impact of nutrient excess, such as palmitate, on circadian rhythms and neuroendocrine signaling pathways, as well as changes in vital neuropeptides, leading to the development of neuronal inflammation; the role of proinflammatory molecules in this process; and ultimately, restoration of normal signaling, clock gene expression, and neuropeptide synthesis in disrupted states by beneficial anti-inflammatory compounds in defined hypothalamic neurons.

Keyword: energy

Binding energies and the entry route of and palmitoylcarnitine into myoglobin.

The interaction of lipids (entry mechanism) with respect to both oxy- and deoxy-myoglobin was explored using unrestrained Molecular Dynamics simulations. The results indicated a spontaneous entry of both and palmitoylcarnitine molecules into the oxy-Mb structure at the main binding site, whereas in deoxy-Mb, both the lipid ligands move away from the protein surface. For the alternative binding locations, entry of the ligands was independent of the oxygenation state. Presented here are the tables with the myoglobin binding energies for and palmitoylcarnitine estimated using Alchemical Free Perturbation approach for the key structures obtained in unrestrained Molecular Dynamics simulations. These data are referenced in the original article "Exploring the entry route of and palmitoylcarnitine into myoglobin", reference number YABBI7787.

Keyword: energy

A promising form-stable phase change material prepared using cost effective pinecone biochar as the matrix of for thermal storage.

A promising new form-stable phase change material (PA/PB) was fabricated using pinecone biochar (PB) as the supporting material of (PA). The biochar of PB with large surface area was produced by forest residue of pinecone, and it was cheap, environment friendly and easy to prepare. The PB was firstly utilized as the supporter of PA and the characterizations of PA/PB were analyzed by the BET, SEM, XRD, DSC, TGA, FT-IR and thermal conductivity tester. The results demonstrated that the PA was physically absorbed by the PB and the crystal structure of the PA was not destroyed. The results of DSC showed that the fusing and crystallization points of the form-stable phase change material with the maximum content of PA (PA/PB-4) were 59.25\u2009°C and 59.13\u2009°C, and its fusing and freezing latent heat were 84.74\u2009kJ/kg and 83.81\u2009kJ/kg, respectively. The results of TGA suggested that the thermal stability of the PA/PB-4 composite was excellent, which could be used for the applications of thermal storage. Furthermore, the thermal conductivity of PA/PB-4 was 0.3926\u2009W/(m∙K), which was increased by 43.76% compared with that of the pure PA. Thus, the study results indicated that the PA/PB-4 had great potential for thermal storage applications.

Keyword: energy

Lupin seed hydrolysate promotes G-protein-coupled receptor, intracellular Ca and enhanced glycolytic metabolism-mediated insulin secretion from BRIN-BD11 pancreatic beta cells.

Lupin seed proteins have been reported to exhibit hypoglycaemic effects in animals and humans following oral administration, however little is known about its mechanism of action. This study investigated the signalling pathway(s) responsible for the insulinotropic effect of the hydrolysate obtained from lupin (Lupinus angustifolius L.) seed extracts utilizing BRIN-BD11 β-cells. The extract was treated with digestive enzymes to give a hydrolysate rich in biomolecules ≤7\u202fkDa. Cells exhibited hydrolysate induced dose-dependent stimulation of insulin secretion and enhanced intracellular Ca and glucose metabolism. The stimulatory effect of the hydrolysate was potentiated by depolarizing concentrations of KCl and was blocked by inhibitors of the ATP sensitive K channel, Gα protein, phospholipase C (PLC) and protein kinase C (PKC). These findings reveal a novel mechanism for lupin hydrolysate stimulated insulin secretion via Gα mediated signal transduction (Gα/PLC/PKC) in the β-cells. Thus, lupin hydrolysates may have potential for nutraceutical treatment in type 2 diabetes.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Association of low oleic intake with diabetic retinopathy in type 2 diabetic patients: a case-control study.

The objective of this study was to describe the intake of macronutrient, especially fatty acids, and explore their possible effect on diabetic retinopathy (DR) in patients with type 2 diabetes mellitus.In this case-control study, we included a total of 146 patients with DR and 148 without DR. The intake of macronutrient was evaluated using a validated food frequency questionnaire. We used logistic regression adjusted for sex, age, diabetes duration, intake, educational level, physical activity, waist circumference, systolic blood pressure, high-density lipoprotein cholesterol and diabetes treatment, to estimate odds ratio (ORs) of DR.Patients with DR had significantly lower intake of fibre, monounsaturated fatty acids (MUFA), and and oleic . Inverse associations were observed between MUFA and oleic intake in DR. Subjects with intermediate and high MUFA intake were less likely to have DR than those with lower MUFA intake, with ORs of 0.46 (95\u2009%\u2009CI: 0.22-0.93) and 0.42 (95\u2009%\u2009CI: 0.18-0.97), respectively. Similarly, intermediate and high oleic intake were associated with reduced DR frequency compared with low oleic intake, with OR values of 0.48 (95\u2009%\u2009CI: 0.23-0.97) and 0.37 (95\u2009%\u2009CI: 0.16-0.85), respectively. These associations were stronger in patients with a longer diabetes duration.In type 2 diabetes mellitus, MUFA and oleic intake were inversely associated with DR.

Keyword: energy

Palmitate Induces an Anti-Inflammatory Response in Immortalized Microglial BV-2 and IMG Cell Lines that Decreases TNFα Levels in mHypoE-46 Hypothalamic Neurons in Co-Culture.

Elevated levels of saturated fatty acids (SFA) induce a state of neuroinflammation in the hypothalamus. It has been suggested that microglia sense palmitate, a prevalent circulating SFA, and act as mediators of this inflammatory process by communicating with neurons, particularly those involved in appetite regulation. In this study, we examined the inflammatory response to palmitate in immortalized microglial cell lines, BV-2 and IMG, and the subsequent effects on inflammatory gene expression in a model of NPY/AgRP neurons, mHypoE-46.The BV-2 cells were treated with 50 µM palmitate for 4 and 24 h, and the transcriptional regulation of markers for inflammation and cellular stress was assessed using an RT2 Profiler PCR Array. Select genes were verified with qRT-PCR. The BV-2 and IMG cells were then co-cultured using 1.0-µm cell culture inserts with an immortalized hypothalamic cell line, mHypoE-46, to investigate potential intercellular communication between microglia and neurons.We found that palmitate increased the mRNA levels of specific inflammatory genes, and a general anti-inflammatory profile was revealed in the microglia cells. The mRNA changes in TNFα at 4 and 24 h in BV-2 cells were abrogated with the toll-like receptor 4 (TLR4) inhibitor, TAK-242, indicating the involvement of TLR4. Co-culture of mHypoE-46 neurons with microglia pre-treated with palmitate resulted in repression of TNFα expression in the hypothalamic neurons. As palmitate significantly increased IL-13 expression in microglia, the effect of this cytokine was tested in mHypoE-46 neurons. The addition of IL-13 to neuronal cultures normalized the palmitate-mediated increase in IL-6 and AgRP expression, suggesting that microglia may protect surrounding neurons, at least in part, through the release of IL-13.These results suggest a potential anti-inflammatory role of microglia towards the palmitate-induced neuroinflammation, and potentially homeostasis, in hypothalamic neurons.© 2018 ©2018S. Karger AG, Basel.

Keyword: energy

Recovery and Utilization of Palm Oil Mill Effluent Source as Value-Added Food Products.

The environmental impacts of palm oil mill effluent (POME) have been a concern due to the water pollution and greenhouse gases emissions. Thus, this study was conducted to recover the value-added products from POME source before being discharged. The samples, before (X) and after (Y) the pre-recovery system in the clarification tank were sampled and analysed and proximate analysis indicated that both samples are rich source of food due to high contents of fats and carbohydrates. GCMS analysis showed that the oil extracts contain predominantly , oleic, linoleic and stearic acids. Regiospecific analysis of oil extracts by quantitative C-NMR spectroscopy demonstrated that both oil extracts contain similar degree of saturation of fatty acids at sn-2 and sn-1,3 positions. The samples are rich in various phytonutrients, pro-vitamin A, vitamin E, squalene and phytosterols, thus contributing to exceptionally high total flavonoid contents and moderate antioxidant activities. Overall, samples X and Y are good alternative food sources, besides reducing the environmental impact of POME.

Keyword: energy

Proteomic effects of repeated-dose oral exposure to 2-monochloropropanediol and its dipalmitate in rat testes.

2- and 3-monochloropropanediol (2-MCPD) and their fatty esters are food contaminants which are concomitantly formed upon thermal treatment of foodstuff containing fats and salt. Exposure to 2- or 3-MCPD thus results, for example, from refined vegetable oils, in instant meals or infant formula, as well as in cereals or pastries. The molecular mechanisms of 2-MCPD toxicity are poorly understood. Here, we performed a comprehensive proteomic analysis of 2-MCDP-induced alterations in the testes from rats following oral administration of 10\u202fmg/kg body weight per day 2-MCPD, or an equimolar dose of 2-MCPD dipalmitate as a representative 2-MCPD fatty ester. In the absence of overt histopathologically detectable toxicity, moderate alterations in cellular proteomic signatures were recorded. The observations are in line with the assumption that the molecular mechanisms of 2-MCPD and 3-MCPD toxicity differ. Observed proteomic alterations point towards effects of 2-MCPD on mitogen-dependent signaling and mitochondrial utilization. Presented data for the first time provide insight into proteomic effects of 2-MCPD in testicular tissue.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Bacterial Microbiota and Metabolic Character of Traditional Sour Cream and Butter in Buryatia, Russia.

Traditional sour cream and butter are widely popular fermented dairy products in Russia for their flavor and nutrition, and contain rich microbial biodiversity, particularly in terms of lactic bacteria (LAB). However, few studies have described the microbial communities and metabolic character of traditional sour cream and butter. The objective of this study was to determine the bacterial microbiota and metabolic character of eight samples collected from herdsmen in Buryatia, Russia. Using single-molecule real-time (SMRT) sequencing techniques, we identified a total of 294 species and/or subspecies in 169 bacterial genera, belonging to 14 phyla. The dominant phylum was Firmicutes (81.47%) and the dominant genus was (59.28%). There were differences between the bacterial compositions of the sour cream and butter samples. The relative abundances of , , and were significantly higher in sour cream than in butter, and the abundance of was significantly lower in sour cream than in butter. Using a pure culture method, 48 strains were isolated and identified to represent seven genera and 15 species and/or subspecies. Among these isolates, subsp. (22.50%) was the dominant LAB species. Ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry at elevated was used in combination with statistical methods to detect metabolite differences between traditional sour cream and butter. A total of 27,822 metabolites were detected in all samples, and Lys-Lys, isohexanal, , Leu-Val, and 2\'-deoxycytidine were the most dominant metabolites found in all samples. In addition, 27 significantly different metabolites were detected between the sour cream and butter samples, including short peptides, organic acids, and amino acids. Based on correlation analyses between the most prevalent bacterial species and the main metabolites in sour cream, we conclude that there may be a connection between the dominant LAB species and these metabolites. This study combined omics techniques to analyze the bacterial diversity and metabolic character of traditional sour cream and butter, and we hope that our findings will enrich species resource libraries and provide valuable resources for further research on dairy product flavor.

Keyword: energy

Toxicological evaluation of 2-dodecylcyclobutanone, a unique radiolytic compound of .

This study was conducted to evaluate the toxic effects and potency of 2-dodecylcyclobutanone (2-dDCB), a unique compound derived from via irradiation. In a series of assays of bacterial reverse-mutation, in vitro chromosomal aberration, and in vivo micronucleus, negative responses were found by the treatment of 2-dDCB comparing vehicle control, dimethyl sulfoxide or corn oil. In the acute oral toxicity test, all of the mice administrated 2-dDCB survived, and there were no clinical and necropsy signs observed at any doses (0, 300, and 2000\u202fmg/kg body weight) during the experimental period of 14 days. These results suggested that 2-dDCB is a relatively non-toxic substance with median lethality dose higher than 2000\u202fmg/kg body weight. Moreover, there were no adverse effects noted in rats orally administrated 2-dDCB everyday via gavage for 28 days, even at the highest dose (2.0\u202fmg/kg body weight/day) tested, which is 1000-times higher than the human daily intake of 2-dDCB estimated through an extreme exposure scenario. Overall, these results indicate that 2-dDCB is not likely to raise any human health concerns and irradiated foods containing can be recognized as safe for human consumption under the current international regulation systems for food irradiation.Copyright © 2018. Published by Elsevier Ltd.

Keyword: energy

Rice bran oil ameliorates inflammatory responses by enhancing mitochondrial respiration in murine macrophages.

Previous studies have revealed the anti-inflammatory properties of rice bran oil (RBO), but the detailed mechanisms are poorly understood. Recent studies on the molecular/cellular anti-inflammatory mechanisms of dietary components have demonstrated that mitochondrial respiration plays a key role in macrophage functioning. Since dietary lipids are major substrates for mitochondrial respiration through β-oxidation, the current study examined whether RBO regulates inflammatory responses by modulating mitochondrial metabolism. Palm oil (PO), enriched with which are known to be effectively taken up by cells and used for oxidative phosphorylation, served as a positive control. In the in vitro model of LPS-stimulated RAW 264.7 murine cells, the levels of pro-inflammatory cytokines (IL-6 and TNF-α) in the culture supernatant were significantly reduced by RBO treatment. In contrast, secretion of the anti-inflammatory cytokine IL-10 was upregulated by RBO. Transcription of genes encoding inflammatory mediator molecules (COX-2 and iNOS) and expression of activation markers (CD80, CD86, and MHC-II) in LPS-stimulated RAW 264.7 cells were suppressed by RBO. Mitochondrial respiration (as assessed by an extracellular flux analyzer) increased upon RBO treatment, as the basal respiration, maximal respiration, ATP production, and spare respiratory capacity were upregulated. In an in vivo study, C57BL/6 mice were fed a negative control diet containing corn oil (CO), PO, or RBO for 4 weeks, and bone marrow-derived macrophages (BMDM) were isolated from their tibias and femurs. In pro-inflammatory M1-polarized BMDM (M1-BMDM), the RBO-induced suppression of IL-6 and TNF-α was recapitulated in vivo. Mitochondrial respiration in M1-BMDM also increased following the RBO intervention and the PO control treatment as compared to CO fed negative control. Overall, the current study for the first time demonstrates that RBO regulates inflammatory responses in murine macrophages by upregulating mitochondrial respiration. Further clinical studies are required to validate the animal study.

Keyword: energy

GC-MS Metabolomics Reveals Distinct Profiles of Low- and High-Grade Bladder Cancer Cultured Cells.

Previous studies have shown that metabolomics can be a useful tool to better understand the mechanisms of carcinogenesis; however, alterations in biochemical pathways that lead to bladder cancer (BC) development have hitherto not been fully investigated. In this study, gas chromatography-mass spectrometry (GC-MS)-based metabolomics was applied to unveil the metabolic alterations between low-grade and high-grade BC cultured cell lines. Multivariable analysis revealed a panel of metabolites responsible for the separation between the two tumorigenic cell lines. Significantly lower levels of fatty acids, including myristic, , and palmitoleic acids, were found in high-grade versus low-grade BC cells. Furthermore, significantly altered levels of some amino acids were observed between low- and high-grade BC, namely glycine, leucine, methionine, valine, and aspartic . This study successfully demonstrated the potential of metabolomic analysis to discriminate BC cells according to tumor aggressiveness. Moreover, these findings suggest that bladder tumorigenic cell lines of different grades disclose distinct metabolic profiles, mainly affecting fatty biosynthesis and amino metabolism to compensate for higher energetic needs.

Keyword: energy

High vulnerability of the heart and liver to 3-hydroxypalmitic -induced disruption of mitochondrial functions in intact cell systems.

Patients affected by long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency predominantly present severe liver and cardiac dysfunction, as well as neurological symptoms during metabolic crises, whose pathogenesis is still poorly known. In this study, we demonstrate for the first time that pathological concentrations of 3-hydroxypalmitic (3HPA), the long-chain hydroxyl fatty (LCHFA) that most accumulates in LCHAD deficiency, significantly decreased adenosine triphosphate-linked and uncoupled mitochondrial respiration in intact cell systems consisting of heart fibers, cardiomyocytes, and hepatocytes, but less intense in diced forebrain. 3HPA also significantly reduced mitochondrial Ca retention capacity and membrane potential in Ca -loaded mitochondria more markedly in the heart and the liver, with mild or no effects in the brain, supporting a higher susceptibility of the heart and the liver to the toxic effects of this fatty . It is postulated that disruption of mitochondrial and Ca homeostasis caused by the accumulation of LCHFA may contribute toward the severe cardiac and hepatic clinical manifestations observed in the affected patients.© 2018 Wiley Periodicals, Inc.

Keyword: energy

Coupling natural systems with synthetic chemistry for light-driven enzymatic biocatalysis.

Visible light-driven redox reactions have been widely adopted for the production of chemicals to combat shortage and global warming. Key elements of such a reaction system include a photosensitizer, a catalyst, and an electron source. In this review, we introduce the small molecules and nanoparticles that are widely used as photosensitizers, as well as the development of a photosensitizer protein that is based on the expansion of genetic code, with a fluorescent protein that is used as a scaffold. Visible light-driven enzymes using proteins as photosensitizers or as catalysts such as carbon monoxide dehydrogenase (CODH), formic dehydrogenase (FDH), hydrogenase, nitrogenase, cytochrome P450 BM3, and alkane synthase are then described. CODH can be coupled with photosensitizing nanoparticles to reduce CO to CO, and hydrogenase can produce H using high- electrons produced from dye-sensitized nanoparticles. When water-soluble zinc porphyrin is coupled with FDH, visible light drives CO to produce formic . Nitrogenase can reduce N to NH using CdS nanoparticle as photosensitizer. Cytochrome P450 BM3 can be enhanced by a visible light-driven redox system and thus by hydroxylate lauric or fatty acids. CvFAP, an alkane synthase, can decarboxylate to pentadecane under blue light excitation. Moreover, we describe a genetically encoded photosensitive protein, which mimics the function of natural photosynthesis and catalyzes the conversion of CO to CO when covalently attached with a Ni-terpyridine complex.

Keyword: energy

Effect of dietary n-3 polyunsaturated fatty supplementation and post-insemination plane of nutrition on systemic concentrations of metabolic analytes, progesterone, hepatic gene expression and embryo development and survival in beef heifers.

Nutrition, and particularly dietary intake, plays a fundamental role in reproductive function in cattle. There is some evidence that supplemental omega-3 dietary polyunsaturated fatty acids (n-3 PUFA) can exert positive effects on fertility. The objectives of this study were to evaluate the effect of dietary n-3 PUFA supplementation, post-insemination plane of nutrition and their interaction on embryo survival in cattle. Crossbred beef heifers (n\u202f=\u202f185) were individually offered barley straw ad libitum and 6\u202fkg DM of concentrate supplemented with either a rumen-protected source of saturated fatty (; control, CON) or a partially rumen-protected n-3 PUFA-enriched supplement (n-3 PUFA). Estrous was synchronised using two injections of PG administered at 11-d intervals and following artificial insemination (AI\u202f=\u202fDay 0) 179 heifers exhibiting oestrus were inseminated and assigned to one of two dietary treatments: (i) remain on their pre-insemination high dietary plane of nutrition (High) or (ii) restricted to 0.6\xa0× estimated maintenance requirements (Low) in a 2\u202f×\u202f2 factorial design. The heifers were then maintained on their assigned diets until slaughter and embryo recovery on Day 16 (n\u202f=\u202f92) or pregnancy diagnosis by ultrasound scanning at Day 30 post-AI (n\u202f=\u202f87). Plasma concentrations of fatty acids, metabolites, insulin, progesterone (P4) and insulin-like growth factor 1 (IGF-1) were measured at appropriate intervals. Hepatic expression of mRNA for aldo-keto reductase (AKR1C), cytochrome P450 2C (CYP 2C) and cytochrome P450 3A (CYP 3A) was examined. The n-3 PUFA supplementation increased plasma n-3 PUFA concentration (P\u202f<\u202f0.05) and reduced n-6: n-3 PUFA ratio (P\u202f<\u202f0.05). Plasma IGF-1 was higher for n-3 PUFA relative to the CON (P\u202f<\u202f0.05) and for High compared with Low plane of nutrition post-AI (P\u202f<\u202f0.05) groups. A low plane of nutrition post-AI increased plasma concentrations of progesterone from Days 7-16 after insemination (P\u202f<\u202f0.001) but reduced embryo length (P\u202f<\u202f0.001). Supplementation with n-3 PUFA reduced and tended to reduce hepatic expression of CYP2C (P\u202f=\u202f0.01) and CYP3A (P\u202f=\u202f0.08), respectively. However, while dietary n-3 PUFA supplementation and an abrupt reduction in nutrient status following insemination elevated plasma concentrations of n-3 PUFA and mid and late phase P4, respectively, there was no effect of either PUFA supplementation or post-insemination plane of nutrition on embryo survival.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: energy

Development of a high-throughput method for real-time assessment of cellular metabolism in intact long skeletal muscle fibre bundles.

We developed a method that allows for real-time assessment of cellular metabolism in isolated, intact long skeletal muscle fibre bundles from adult mice. This method can be used to study changes in mitochondrial function and fuel utilisation in live skeletal muscle fibre bundles. Our method enables flexibility in experimental design and high-throughput assessment of mitochondrial parameters in isolated skeletal muscle fibre bundles. Extensor digitorum longus (EDL) fibre bundles obtained from chronic high-fat diet fed mice had lower basal oxygen consumption under FCCP-induced maximal respiration, when compared to control chow-fed mice. EDL fibre bundles obtained from chronic high-fat diet fed mice had enhanced mitochondrial oxidation capacity under FCCP-induced maximal respiration, when compared to control chow-fed mice.Metabolic dysfunction in skeletal muscle contributes to the aetiology and development of muscle diseases and metabolic diseases. As such, assessment of skeletal muscle cellular bioenergetics provides a powerful means to understand the role of skeletal muscle metabolism in disease and to identify possible therapeutic targets. Here, we developed a method that allows for the real-time assessment of cellular respiration in intact skeletal muscle fibre bundles obtained from the extensor digitorum longus (EDL) muscle of adult mice. Using this method, we assessed the contribution of ATP turnover and proton leak to basal mitochondrial oxygen consumption rate (OCR). Our data demonstrate that the mitochondria in EDL fibres are loosely coupled. Moreover, in the presence of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), we show that palmitate exposure induced comparable peak OCR and higher total OCR in EDL fibre bundles when compared to pyruvate exposure, suggesting that fatty acids might be a more sustainable fuel source for skeletal muscle when mitochondria are driven to maximal respiration. Application of this method to EDL fibre bundles obtained from chronic high-fat diet fed mice revealed lower basal OCR and enhanced mitochondrial oxidation capacity in the presence of FCCP when compared to the chow-diet fed control mice. By using a 96-well microplate format, our method provides a flexible and efficient platform to investigate mitochondrial parameters of intact skeletal muscle fibres obtained from adult mice.© 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.

Keyword: energy

Comparative analysis of nutrient content and of eggs from different chicken genotypes.

Eggs are important foods in the daily diet of humans and have great biological activity and a high digestibility. Egg yolk is a good source of biologically active substances such as fatty acids, phospholipids, sterols and tocopherols. The eggs of seven chicken genotypes were analyzed for their chemical composition, and a detailed study of the lipids in egg yolk was conducted. composition of the egg yolk and egg albumen was 29.06-30.51\u2009MJ\u2009kg and 19.77-20.93\u2009MJ\u2009kg respectively. Regarding their chemical composition: water ranged from 471.7 to 515.4\xa0g\u2009kg and 878.3-885.9\xa0g\u2009kg ; fat content in dry matter ranged from 607 to 647\u2009g\u2009kg and 6.7-11.6\xa0g\u2009kg ; protein varied from 302 to 331.7\xa0g\u2009kg and 823.6-892.5\xa0g\u2009kg ; ash ranged from 33.7 to 37.7\xa0g\u2009kg and 63.8-74.0\xa0g\u2009kg ; and nitrogen-free extracts ranged from 12.7 to 36.5\xa0g\u2009kg and 35.0-96.2\xa0g\u2009kg . The sterols and phospholipids in the yolk lipids were 16-26\u2009g\u2009kg and 59-127\u2009g\u2009kg . The main fatty acids in the lipids were oleic (39.1-47.3%) and (26.0-35.5%) acids. Cholesterol in the yolk lipids ranged from 15.9 to 25.9\xa0g\u2009kg . Phosphatidylcholine (389-573\u2009g\u2009kg ), phosphatidylethanolamine (219-355\u2009g\u2009kg ) and phosphatidylinositol (112-284\u2009g\u2009kg ) were the main phospholipids. The content of saturated fatty acids in the phospholipids was significantly higher than that in triacylglycerols.Small variations in the chemical composition of eggs from seven different genotypes were observed. Significant differences in the fatty compositions of the main classes of phospholipids and the triacylglycerol fraction were established. © 2019 Society of Chemical Industry.© 2019 Society of Chemical Industry.

Keyword: energy

The protein-sparing effect of α-lipoic in juvenile grass carp, Ctenopharyngodon idellus: effects on lipolysis, fatty β-oxidation and protein synthesis.

To investigate the protein-sparing effect of α-lipoic (LA), experimental fish (initial body weight: 18·99 (sd 1·82) g) were fed on a 0, 600 or 1200 mg/kg α-LA diet for 56 d, and hepatocytes were treated with 20 μm compound C, the inhibitor of AMP kinase α (AMPKα), treated for 30 min before α-LA treatment for 24 h. LA significantly decreased lipid content of the whole body and other tissues (P0·05). Consistent with results from the experiment in vitro, LA activated phosphorylation of AMPKα and notably increased the protein content of adipose TAG lipase in intraperitoneal fat, hepatopancreas and muscle in vivo (P<0·05). Meanwhile, LA significantly up-regulated the mRNA expression of genes involved in fatty β-oxidation in the same three areas, and LA also obviously down-regulated the mRNA expression of genes involved in amino catabolism in muscle (P<0·05). Besides, it was observed that LA significantly activated the mammalian target of rapamycin (mTOR) pathway in muscle of experimental fish (P<0·05). LA could promote lipolysis and fatty β-oxidation via increasing supply from lipid catabolism, and then, it could economise on the protein from production to increase protein deposition in grass carp. Besides, LA might directly promote protein synthesis through activating the mTOR pathway.

Keyword: energy

Recycling of food waste for fuel precursors using an integrated bio-refinery approach.

The main aim was to integrate FW-recycling with cultivation of Rhodotorula glutinis and anaerobic digestion (AD) for bio- and -fuel recovery. Mixed FW was mechanically macerated (Pcon) and hydrolysed (at 250gL water) via chemical (Ch), thermal (Th) and TCh (combined Ch and Th) treatments. Cleared hydrolysates from individual pre-treatment processes were used as culture medium for cultivation of R. glutinis, while the residual solids (RS) were subjected to AD. Pcon cultivation yielded maximal R. glutinis dry weight biomass (5.18gL) and total fatty contents (1.03gg DW). Dominant fatty methyl esters (FAME) were - (C-26%); Stearic - (C-17%) and Oleic acids (C-38%), ideal for bio-diesel production. Highest methane yields (actual ∼0.139mkg volatile solids) were measured from AD of Th-derived RS. Thus integrated FW recycling approaches will be more feasible for generating and economic incentives.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Effects of Stanniocalcin-1 on glucose flux in rat brown adipose tissue.

The present work assesses in\xa0vitro the role of human Stanniocalcin 1 (hSTC-1) in C-glucose metabolism in brown adipose tissue (BAT) from fed rat. In the fed state, hSTC-1 decreases the incorporation of C from glucose into lipids in the rat BAT. The data support the hypothesis that the capacity of the glycerol-3-phosphate (G3P)-generating pathway (glycolysis) from glucose is regulated by hSTC-1, decreasing the adequate supply of G3P needed for fatty esterification and triacylglycerol (TG) storage in BAT. The results also suggest the effect of hSTC-1 on de novo fatty synthesis from pyruvate generated by C-glucose in the glycolysis pathway. In addition, by decreasing lipogenesis, hSTC-1 increased ATP levels and these two factors may decrease BAT thermogenic function. The presence of hSTC-1 in the incubation medium did not alter C-glucose and C-1- oxidation. The uncoupling protein 1 (UCP-1) expression was not altered by hSTC-1 either. In conclusion, hSTC-1 is one of the hormonal factors that control glucose metabolism in BAT in the fed state. The decrease of TG capacity synthesis from C-glucose by hSTC-1 compromises the BAT thermogenic capacity. Furthermore, the increase in ATP levels would inhibit a futile cycle via UCP-1, which dissipates oxidative as heat.Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Keyword: energy

The digestible , metabolizable , and net content of dietary fat sources in thirteen- and fifty-kilogram pigs.

The objective was to determine the concentration of a diverse array of dietary fat sources and, from these data, develop regression equations that explain differences based on chemical composition. A total of 120 Genetiporc 6.0 × Genetiporc F25 (PIC, Inc., Hendersonville, TN) individually housed barrows were studied for 56 d. These barrows (initial BW of 9.9 ± 0.6 kg) were randomly allotted to 1 of 15 dietary treatments. Each experimental diet included 95% of a corn-soybean meal basal diet plus 5% either corn starch or 1 of 14 dietary fat sources. The 14 dietary fat sources (animal-vegetable blend, canola oil, choice white grease source A, choice white grease source B, coconut oil, corn oil source A, corn oil source B, fish oil, flaxseed oil, palm oil, poultry fat, soybean oil source A, soybean oil source B, and tallow) were selected to provide a diverse and robust range of unsaturated fatty :SFA ratios (U:S). Pigs were limit-fed experimental diets from d 0 to 10 and from d 46 to 56, providing a 7-d adaption for fecal collection on d 7 to 10 (13 kg BW) and d 53 to 56 (50 kg BW). At 13 kg BW, the average content of the 14 sources was 8.42 Mcal DE/kg, 8.26 Mcal ME/kg, and 7.27 Mcal NE/kg. At 50 kg BW, the average content was 8.45 Mcal DE/kg, 8.28 Mcal ME/kg, and 7.29 Mcal NE/kg. At 13 kg BW, the variation of dietary fat DE content was explained by DE (Mcal/kg) = 9.363 + [0.097 × (FFA, %)] - [0.016 × omega-6:omega-3 fatty acids ratio] - [1.240 × (arachidic , %)] - [5.054 × (insoluble impurities, %)] + [0.014 × (, %)] ( = 0.008, = 0.82). At 50 kg BW, the variation of dietary fat DE content was explained by DE (Mcal/kg) = 8.357 + [0.189 × U:S] - [0.195 × (FFA, %)] - [6.768 × (behenic , %)] + [0.024 × (PUFA, %)] ( = 0.002, = 0.81). In summary, the chemical composition of dietary fat explained a large degree of the variation observed in the content of dietary fat sources at both 13 and 50 kg BW.

Keyword: energy

Increased triacylglycerol - Fatty substrate cycling in human skeletal muscle cells exposed to eicosapentaenoic .

It has previously been shown that pretreatment of differentiated human skeletal muscle cells (myotubes) with eicosapentaenoic (EPA) promoted increased uptake of fatty acids and increased triacylglycerol accumulation, compared to pretreatment with oleic (OA) and (PA). The aim of the present study was to examine whether EPA could affect substrate cycling in human skeletal muscle cells by altering lipolysis rate of intracellular TAG and re-esterification of fatty acids. Fatty metabolism was studied in human myotubes using a mixture of fatty acids, consisting of radiolabelled oleic as tracer (14C-OA) together with EPA or PA. Co-incubation of myotubes with EPA increased cell-accumulation and incomplete fatty oxidation of 14C-OA compared to co-incubation with PA. Lipid distribution showed higher incorporation of 14C-OA into all cellular lipids after co-incubation with EPA relative to PA, with most markedly increases (3 to 4-fold) for diacylglycerol and triacylglycerol. Further, the increases in cellular lipids after co-incubation with EPA were accompanied by higher lipolysis and fatty re-esterification rate. Correspondingly, basal respiration, proton leak and maximal respiration were significantly increased in cells exposed to EPA compared to PA. Microarray and Gene Ontology (GO) enrichment analysis showed that EPA, related to PA, significantly changed i.e. the GO terms "Neutral lipid metabolic process" and "Regulation of lipid storage". Finally, an inhibitor of diacylglycerol acyltransferase 1 decreased the effect of EPA to promote fatty accumulation. In conclusion, incubation of human myotubes with EPA, compared to PA, increased processes of fatty turnover and oxidation suggesting that EPA may activate futile substrate cycling of fatty acids in human myotubes. Increased TAG-FA cycling may be involved in the potentially favourable effects of long-chain polyunsaturated n-3 fatty acids on skeletal muscle and whole-body metabolism.

Keyword: energy

Effect of co-culture with Tetragenococcus halophilus on the physiological characterization and transcription profiling of Zygosaccharomyces rouxii.

Zygosaccharomyces rouxii and Tetragenococcus halophilus are widely existed and play vital roles during the manufacture of fermented foods such as soy sauce. The aim of this study was to elucidate the effect of T. halophilus CGMCC 3792 on the physiological characterizations and transcription profiling of Z. rouxii CGMCC 3791. Salt tolerance analysis revealed that co-culture with T. halophilus enhanced the salt tolerance of Z. rouxii during salt stress. Analysis of the volatile compounds revealed that co-culture reduced the level of 1-butanol, improved the level of octanoic which all were produced by T. halophilus and reduced the level of phenylethyl alcohol produced by Z. rouxii. The presence of Z. rouxii decreased the contents of 3,4-dimethylbenzaldehyde and acetic produced by T. halophilus. In addition, co-culture improved the content of benzyl alcohol significantly. Analysis of membrane fatty showed that co-culture improved the content of (C16:0) and stearic (C18:0) acids in cells of Z. rouxii, and reduced the contents of myristic (C14:0), palmitoleic (C16:1) and oleic (C18:1). In order to further explore the interactions between the two strains, RNA-seq technology was used to investigate the effect of co-culture with T. halophilus on the transcription profiling of Z. rouxii. By comparing cells incubated in co-culture group with cells incubated in single-culture group, a total of 967 genes were considered as differentially expressed genes (DEGs). Among the DEGs, 72 genes were up-regulated, while 895 genes were down-regulated. These DEGs took party in various activities in cells of Z. rouxii, and the result showed co-culture with T. halophilus had a positive effect on proteolysis, the attachment of a cell to another cell, extracellular protein accumulation, metabolism, and a negative effect on oxidative phosphorylation, small molecular substances metabolism, DNA replication and repair, and transcription in cells of Z. rouxii. Results presented in this study may contribute to further understand the interactions between Zygosaccharomyces rouxii and Tetragenococcus halophilus.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: energy

[Insulin resistance is an alimentary deficiency of substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: energy

Prolyl Oligopeptidase Inhibition Attenuates Steatosis in the L02 Human Liver Cell Line.

Prolyl oligopeptidase (POP) is a serine endopeptidase that is widely distributed in vivo, particularly in the liver. Significant changes in functional mitochondrial proteins involved with mitochondrial oxidoreductases/transporters and nucleic binding proteins were observed after POP inhibition in the liver, which suggested a role of POP in regulating liver metabolism. Steatosis in nonalcoholic fatty liver disease (NAFLD) is associated with disturbances in lipid and metabolism in hepatocytes. Here, we aimed to study the effect of POP on hepatocyte steatosis.The human liver cell line L02 was used to investigate the biological effects of POP. An in vitro cell model of steatosis was successfully induced with oleic and . L02 cells were also subjected to S17092 (a POP inhibitor) at different concentrations for 24 or 48 h. Ac-SDKP levels and POP activity were measured to assess the rate of inhibition of POP by S17092. The POP gene and protein expression levels were detected using real-time PCR and Western blots, respectively. Oil red O staining was performed and the triglyceride levels in the L02 cells were also measured. Cell proliferation and apoptosis were detected using CCK-8 and flow cytometry, respectively. The expression of genes involved in lipid metabolism was detected using real-time PCR. The effects of POP inhibition on LC3B II were detected by Western blot.Compared with the control, the POP mRNA levels increased by approximately 30%, and the POP protein levels increased by almost 60% in the steatotic L02 cells. After S17092 (0.026~130 μM) incubation for 24 or 48 h, cell proliferation was significantly decreased in the free fatty (FFA)-treated cells at 26-130 μM; however, S17092 did not affect the proliferation of L02 cells after 24 h of incubation with S17092 at 0.026-65 μM without FFA treatment. S17092 treatment (13 and 26 μM) also elicited no significant effect on apoptosis in normal L02 cells, but FFA treatment increased cell apoptosis, which was attenuated by S17092 incubation. S17092 treatment inhibited intracellular POP activity and decreased the AcSDKP level at the concentration of 0.026-26 μM. After treatment with FFA for 24 h, oil red O staining revealed significant lipid accumulation in the cells in the model group compared with the controls; however, lipid accumulation was suppressed after the administration of S17092 (13 and 26 μM). Accordingly, the triglyceride levels in the FFA-treated cells were approximately 5-fold greater than those of the controls and were decreased by approximately 25% and 45% after the administration of S17092 at 13 and 26 μM, respectively. The mRNA levels of FASN, PPAR-γ, and SREBP-1c were higher in the FFA-treated cells than in the normal controls, and all of these levels were significantly inhibited in the presence of S17092 at both 13 and 26 μM. S17092 treatment did not affect LC3B II in the FFA-treated cells compared with FFA treatment alone.The expression of POP increases with hepatocyte steatosis, and POP inhibitors can significantly reduce intracellular lipid accumulation, which might be related to the inhibition of genes involved in lipid synthesis.

Keyword: energy

Acute hypoxic preconditioning prevents -induced cardiomyocyte apoptosis via switching metabolic GLUT4-glucose pathway back to CD36-fatty dependent.

Metabolic syndrome is a risk factor for the development of cardiovascular diseases. Myocardial cell damage leads to an imbalance of metabolism, and many studies have indicated that short-term hypoxia during myocardial cell injury has a protective effect. In our previous animal studies, we found that short-term hypoxia in the heart has a protective effect, but long-term hypoxia increases myocardial cell injury. (PA)-treated H9c2 cardiomyoblasts and neonatal rat ventricle cardiomyocytes were used to simulate hyperlipidemia model, which suppress cluster of differentiation 36 (CD36) and activate glucose transporter type 4 (GLUT4). We exposed the cells to short- and long-term hypoxia and investigated the protective effects of hypoxic preconditioning on PA-induced lipotoxicity in H9c2 cardiomyoblasts and neonatal rat cardiomyocytes. Preconditioning with short-term hypoxia enhanced both CD36 and GLUT4 metabolism pathway protein levels. Expression levels of phospho-PI3K, phospho-Akt, phospho-AMPK, SIRT1, PGC1α, PPARα, CD36, and CPT1β induced by PA was reversed by short-term hypoxia in a time-dependent manner. PA-induced increased GLUT4 membrane protein level was reduced in the cells exposed to short-term hypoxia and si-PKCζ. Short-term hypoxia, resveratrol and si-PKCζ rescue H9c2 cells from apoptosis induced by PA and switch the metabolic pathway from GLUT4 dependent to CD36 dependent. We demonstrate short-term hypoxic preconditioning as a more efficient way as resveratrol in maintaining the metabolism of hearts during hyperlipidemia and can be used as a therapeutic strategy.© 2017 Wiley Periodicals, Inc.

Keyword: energy

Potential of Whole Pequi ( spp.) Fruit-Pulp, Almond, Oil, and Shell-as a Medicinal Food.

Pequi () pulp, the most consumed component of pequi fruit, is one of the richest Brazilian carotenoid sources, and the most important carotenoid food source native to the Cerrado. However, there are considerable differences among pequi species regarding total carotenoids content and carotenoids profile. Camb. pulp presents higher content of total carotenoids than (Aubl.) Pers. Regarding the carotenoids profile, few studies are available in the literature, mainly with Pequi pulp also has high contents of lipids, dietary fiber, zinc, and magnesium, and is source of calcium and polyphenols. Pequi almond presents high , lipid, protein, dietary fiber, and ash contents. Pequi oil (pulp and almond) has high levels of monounsaturated fatty acids, especially oleic , and relatively high contents of saturated fatty acids, mainly . Pequi shell (exocarp and external mesocarp) is the largest component of the fruit and a solid residue of the pequi processing, which is rich in dietary fibers, including soluble fibers, and phenolic compounds, mostly gallic , ellagic , and quercetin. Pulp oil is the pequi byproduct most investigated in studies. Research with pequi pulp oil in animal models has shown antioxidant, anti-inflammatory, cardioprotective, hepatoprotective, antigenotoxic, and anticarcinogenic effects. In humans, there are evidences supporting anti-inflammatory, cardioprotective, and antigenotoxic effects. Studies on carotenoids profile of pequi pulp in different fruit species are recommended, and studies are necessary to better explore the potential health benefits of pequi fruit components, mainly the pequi pulp and shell.

Keyword: energy

Palm oil and human health. Meeting report of NFI: Nutrition Foundation of Italy symposium.

The use of palm oil by the food industry is increasingly criticized, especially in Italy, for its purported negative effects on human health and environment. This paper summarizes the conclusions of a Symposium on this topic, gathered by the Nutrition Foundation of Italy, among experts representing a number of Italian Medical and Nutritional Scientific Societies. Toxicological and environmental issues were not considered. Participants agreed that: no evidence does exist on the specific health effects of palm oil consumption as compared to other saturated fatty acids-rich fats; the stereospecific distribution of saturated fatty acids in the triacylglycerol molecule of palm oil limits their absorption rate and metabolic effects; in agreement with International guidelines, saturated fatty acids intake should be kept\u2009<10% of total , within a balanced diet; within these limits, no effect of palm oil consumption on human health (and specifically on CVD or cancer risk) can be foreseen.

Keyword: energy

Effect of conjugated linoleic and acetate on milk fat synthesis and adipose lipogenesis in lactating dairy cows.

During biohydrogenation-induced milk fat depression (MFD), nutrients are spared from milk fat synthesis and are available for other metabolic uses. Acetate is the major carbon source spared and it may increase lipid synthesis in adipose tissue during MFD. The objective of this study was to compare the effect of trans-10,cis-12 conjugated linoleic (CLA) and the amount of acetate spared during CLA-induced MFD on adipose tissue lipogenesis. Nine multiparous, lactating, ruminally cannulated Holstein cows (244 ± 107 d in milk; 25 ± 8.4 kg of milk/d; mean ± standard deviation) were randomly assigned to treatments in a 3 × 3 Latin square design. Experimental periods were 4 d followed by a 10-d washout. Treatments were control (CON), ruminal infusion of acetate (AC; continuous infusion of 7 mol/d adjusted to pH 6.1 with sodium hydroxide), or abomasal infusion of CLA (10 g/d of both trans-10,cis-12 CLA and cis-9,trans-11 CLA). Dry matter intake, milk yield, and milk protein yield and percentage were not affected by treatments. Compared with CON, milk fat yield decreased 23% and fat percent decreased 28% in CLA, and milk fat yield increased 20% in AC. Concentration and yield of milk de novo synthesized fatty acids (C16) was increased by CLA, compared with CON. Yield of de novo synthesized fatty acids and was increased by AC, compared with CON. Lipogenesis capacity of adipose tissue explants was decreased 72% by CLA, but was not affected by AC. Acetate oxidation by adipose explants was not affected by treatments. Treatments had no effect on expression of key lipogenic factors, lipogenic enzymes, and leptin; however, expression of fatty binding protein 4 was reduced in CLA compared with CON. Additionally, hormone-sensitive lipase and perilipin 1 were decreased by CLA and acetate. Plasma glucose and glucagon concentrations were not affected by treatments; however, CLA increased nonesterified fatty acids 17.7%, β-hydroxybutyrate 16.1%, and insulin 27.8% compared with CON, and AC increased plasma β-hydroxybutyrate 18%. In conclusion, during CLA-induced MFD in low-producing cow adipose tissue was sensitive to the anti-lipogenic effects of CLA, while spared acetate did not stimulate adipose lipogenesis. However, acetate may play an important role in stimulating lipogenesis and improving status in the mammary gland under normal conditions.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Selection and Characterization of Responsive Patients with an OXPHOS Complex I Defect.

Mitochondrial disorders are genetically and clinically heterogeneous, mainly affecting high -demanding organs due to impaired oxidative phosphorylation (OXPHOS). Currently, effective treatments for OXPHOS defects, with complex I deficiency being the most prevalent, are not available. Yet, clinical practice has shown that some complex I deficient patients benefit from a high-fat or ketogenic diet, but it is unclear how these therapeutic diets influence mitochondrial function and more importantly, which complex I patients could benefit from such treatment. Dietary studies in a complex I deficient patient with exercise intolerance showed increased muscle endurance on a high-fat diet compared to a high-carbohydrate diet. We performed whole-exome sequencing to characterize the genetic defect. A pathogenic homozygous p.G212V missense mutation was identified in the gene, encoding an early assembly factor of complex I. A complementation study in fibroblasts confirmed that the p.G212V mutation caused the complex I deficiency. The mechanism turned out to be an incomplete assembly of the peripheral arm of complex I, leading to a decrease in the amount of mature complex I. The patient clinically improved on a high-fat diet, which was supported by the 25% increase in maximal OXPHOS capacity in TMEM126B defective fibroblast by the saturated fatty , whereas oleic did not have any effect in those fibroblasts. Fibroblasts of other patients with a characterized complex I gene defect were tested in the same way. Patient fibroblasts with complex I defects in NDUFS7 and NDUFAF5 responded to , whereas ACAD9, NDUFA12, and NDUFV2 defects were non-responding. Although the data are too limited to draw a definite conclusion on the mechanism, there is a tendency that protein defects involved in early assembly complexes, improve with , whereas proteins defects involved in late assembly, do not. Our data show at a clinical and biochemical level that a high fat diet can be beneficial for complex I patients and that our cell line assay will be an easy tool for the selection of patients, who might potentially benefit from this therapeutic diet.

Keyword: energy

Serum metabolomics study of nutrient metabolic variations in chronic heat-stressed broilers.

To investigate the effects of heat stress on broiler metabolism, we assigned 144 broilers to normal control (NC), heat stress (HS) or pair-fed (PF) groups and then monitored the effects using growth performance, carcass characteristics, biochemical assays and GC-MS-based metabolomics. The up-regulation of cloacal temperature confirmed that our experiment was successful in inducing chronic heat stress. The average daily gain and average daily feed intake of the HS group were significantly lower than those of the NC group, by 28·76 and 18·42 %, respectively (P1 and P<0·05). The greater feed:gain ratio of the HS group was significantly positively correlated with the leg, abdominal fat, subcutaneous fat and intramuscular fat proportions and levels of some free amino acids (proline, l-cysteine, methionine and threonine) but was negatively correlated with breast proportion and levels of some NEFA (stearic , arachidonic , and oleic ). These findings indicated that the heat-stressed broilers were in negative balance and unable to effectively mobilise fat, thereby resulting in protein decomposition, which subsequently affected growth performance and carcass characteristics.

Keyword: energy

Understanding the potency of fatty acids with the amino side chains of bovine β lactoglobulin-A quantum chemical approach.

The present study aims to spotlight on the strength of interaction between different fatty acids with the Bovine-lactoglobulin (LGB) protein side chains along with the crystal water molecules at M062X/def2-QZVP level of theory. To analyse the role of carboxylic and its interaction with side chains and to reveal the significance of carboxylic , it was replaced by Acyl chloride (COCl), Acyl Bromide (COBr) and Acyl-Fluorine (COF) group and COS group. The ligands are linear with a straight and branched chain of carbon atoms, but extended methyl group make the ligand bend resulting in non-planar geometry. The least and highest band gap reveals the conductivity properties of ligands. 3UEW, the is well-built owing to the interaction with the amino side chains Lys 69 and Glu 62 resulting in interaction of -124.98kcal/mol. 3D-NCIplot isosurface map and 2D-NCIplot graph plays a key role to confirm and analyse the occurrence of various non covalent interactions. The overall analysis of the fatty acids implies the fact that depending on the aliphatic chain length, the carboxyl group was capable of positioning favourable binding site.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

Co-overexpression of native phospholipid-biosynthetic genes plsX and plsC enhances lipid production in Synechocystis sp. PCC 6803.

The overexpression of native plsX and plsC genes involving in fatty /phospholipid synthesis first timely-reported the significantly enhanced lipid contents in Synechocystis sp. PCC 6803. Growth rate, intracellular pigment contents including chlorophyll a and carotenoids, and oxygen evolution rate of all overexpressing (OX) strains were normally similar as wild type. For fatty compositions, saturated fatty , in particular (16:0) was dominantly increased in OX strains whereas slight increases of unsaturated fatty acids were observed, specifically linoleic (18:2) and alpha-linolenic (18:3). The plsC/plsX-overexpressing (OX\u2009+\u2009XC) strain produced high lipid content of about 24.3%w/dcw under normal condition and was further enhanced up to 39.1%w/dcw by acetate induction. This OX\u2009+\u2009XC engineered strain was capable of decreasing phaA transcript level which related to poly-3-hydroxybutyrate (PHB) synthesis under acetate treatment. Moreover, the expression level of gene transcripts revealed that the plsX- and plsC/plsX-overexpression strains had also increased accA transcript amounts which involved in the irreversible carboxylation of acetyl-CoA to malonyl-CoA. Altogether, these overexpressing strains significantly augmented higher lipid contents when compared to wild type by partly overcoming the limitation of lipid production.

Keyword: energy

GLP-1 Elicits an Intrinsic Gut-Liver Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, hyperlipidemia, and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN (fatty synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free fatty , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut-liver regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic metabolism.© 2017 American Heart Association, Inc.

Keyword: energy

Effects of a hypoenergetic diet rich in α-linolenic on fatty composition of serum phospholipids in overweight and obese patients with metabolic syndrome.

Plant-derived α-linolenic (ALA) may exert cardioprotective effects. Dietary ALA can undergo desaturation and elongation to form long-chain ω-3 polyunsaturated fatty acids, but the extent to which this occurs in humans is unclear. The aim of the study was to examine the effects of an -restricted diet enriched with ALA on fatty composition of serum phospholipids in patients with metabolic syndrome.The present analysis compared the effects of a hypoenergetic diet high in ALA (3.4\u2009g/d) with a control diet low in ALA (0.9\u2009g/d) on fatty composition of serum phospholipids in 81 overweight or obese patients with features of metabolic syndrome.After a 26-wk intervention, concentration of ALA in serum phospholipids remained constant in both diet groups. The control group had a significant decrease in serum phospholipid eicosapentaenoic concentration, although no significant intergroup difference was observed. Serum phospholipid docosahexaenoic concentration significantly decreased to a similar extent with both interventions. Additionally, both interventions significantly decreased serum phospholipid concentrations of , stearic , total saturated fatty acids, linoleic , total ω-6 and ω-3 polyunsaturated fatty acids, with no effect of diet group on these changes. Compared with the ALA diet, the control diet led to a significant increase in serum phospholipid oleic concentration.Daily intake of 3.4\u2009g of ALA during a 26-wk -restricted diet did not lead to an enrichment of serum phospholipids with ALA and did not increase eicosapentaenoic due to conversion. Additionally, dietary ALA was unable to compensate for a decrease in serum phospholipid docosahexaenoic .Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

GLP-1 Analogue, Exendin-4, Modulates MAPKs Activity but not the Heat Shock Response in Human HepG2 Cells.

Glucagon-like peptide-1 (GLP-1) analogues reduce ER stress and inflammation in key metabolic organs, including the liver. However, their effects on heat shock response (HSR) and mitogen-activated protein kinases (MAPKs) have not yet been elucidated. In the present study, we investigate whether the GLP-1 analogue, exendin-4, triggers the expression of HSR and increases MAPK activity under metabolic stress.The effects of exendin-4 in the presence or absence of (PA; 400 μm) or glucose (30 mm) in the HepG2 liver cell line are assessed using Western blots, quantitative real-time PCR, and label-free proteomics.Heat shock proteins (HSP60, HSP72, HSP90, and GRP78) and other chaperones are not significantly affected by exendin-4 under the conditions tested. In contrast, the presence of exendin-4 alone increases the MAPK phosphorylation levels (JNK, ERK1/2, and p38). For short incubation periods, in the presence of PA or glucose, treatment with exendin-4 exhibits limited effects but significantly attenuates MAPK phosphorylation after a 24-h incubation. Interestingly, canonical signaling pathways, such as EIF2, ILK, PKA, and Rho, are modulated by exendin-4.Identifying new pathways modulated by GLP-1 analogues will provide further insights into their benefits beyond their currently recognized roles in glycemic control, such as MAPK activity, homeostasis, and body weight decrease.© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: energy

The influence of decreased hydraulic retention time on the performance and stability of co-digestion of sewage sludge with grease trap sludge and organic fraction of municipal waste.

The effect of hydraulic retention time ranging from 12 to 20\xa0d on process performance and stability was investigated in two anaerobic completely stirred tank reactors with a working liquid volume equal to 6 litres. The reactors were fed with mixtures containing (on volatile solids basis): 40% of sewage sludge, 30% of organic fraction of municipal waste and 30% of grease trap sludge. The change of hydraulic retention time did not significantly affect process stability. However, methane yields as well as volatile solids removal decreased from 0.54 to 0.47\xa0l per kg of added volatile solids and 65% to 60% respectively, with the decrease of hydraulic retention time. Despite the fact that the best process performance was achieved for hydraulic retention time of 20 days, the obtained results showed that it is also possible to carry out the co-digestion process at shorter hydraulic retention times with good results. Furthermore, gas production rate as well as biogas production at the shortest hydraulic retention time were approximately 46% higher in comparison to results obtained at the longest hydraulic retention time. In this context, the proposed solution seems to be an interesting option, because it provides an unique opportunity for wastewater treatment plants to improve their profitability by enhancing recovery from sludge as well as full utilisation of the existing infrastructure and hence creates a new potential place for alternative treatment of organic industrial waste such as: fat-rich materials or food waste. However, implementation of the solution at wastewater treatment plants is still a big challenge and needs studies including identification of optimal digesting conditions, information about substrate pumping, inhibition thresholds and processing properties. Additionally, due to the characteristics of both co-substrates their introduction to the full-scale digester should be carefully planned due to a potential risk of overloading of the digester. For this reason, a gradual increase of the share of these wastes in the co-digestion mixture is highly recommended, because it will allow for the acclimatization of bacteria as well as prevent overloading. The results of this study show the importance of gradual acclimatization of microorganisms to the changing environmental conditions. It was found that concentration of long chain fatty acids in effluents increased with the reduction of hydraulic retention time, but this phenomenon did not significantly influence the performance and stability of the process probably due to changes hydraulic retention time being gradual. Although for a moderate negative correlation with volatile solids removal was observed.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

GC-MS-Based metabolomics discovers a shared serum metabolic characteristic among three types of epileptic seizures.

Understanding the overall and common metabolic changes of seizures can provide novel clues for their control and prevention. Here, we aim to investigate the global metabolic feature of serum for three types of seizures.We recruited 27 patients who had experienced a seizure within 48h (including 11 who had a generalized seizure, nine who had a generalized seizure secondary to partial seizure and seven who had a partial seizure) and 23 healthy controls. We analyzed the global metabolic changes of serum after seizures using gas chromatography-mass spectrometry-based metabolomics. Based on differential metabolites, the metabolic pathways and their potential to diagnose seizures were analyzed, and metabolic differences among three types of seizures were compared.The metabolic profiles of serum were distinctive between the seizure group and the controls but were not different among the three types of seizures. Compared to the controls, patients with seizures had higher levels of lactate, butanoic , proline and glutamate and lower levels of , linoleic , elaidic , trans-13-octadecenoic , stearic , citrate, cysteine, glutamine, asparagine, and glyceraldehyde in the serum. Furthermore, these differential metabolites had common change trends among the three types of seizures. Related pathophysiological processes reflected by these metabolites are deficit, inflammation, nervous excitation and neurotoxicity. Importantly, transamination inhibition is suspected to occur in seizures. Lactate, glyceraldehyde and trans-13-octadecenoic in serum jointly enabled a precision of 92.9% for diagnosing seizures.There is a common metabolic feature in three types of seizures. Lactate, glyceraldehyde and trans-13-octadecenoic levels jointly enable high-precision seizure diagnosis.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: energy

feeding increases ceramide supply in association with increased milk yield, circulating nonesterified fatty acids, and adipose tissue responsiveness to a glucose challenge.

Reduced insulin action is a key adaptation that facilitates glucose partitioning to the mammary gland for milk synthesis and enhances adipose tissue lipolysis during early lactation. The progressive recovery of insulin sensitivity as cows advance toward late lactation is accompanied by reductions in circulating nonesterified fatty acids (NEFA) and milk yield. Because can promote insulin resistance in monogastrics through sphingolipid ceramide-dependent mechanisms, (C16:0) feeding may enhance milk production by restoring homeorhetic responses. We hypothesized that feeding C16:0 to mid-lactation cows would enhance ceramide supply and ceramide would be positively associated with milk yield. Twenty multiparous mid-lactation Holstein cows were enrolled in a study consisting of a 5-d covariate, 49-d treatment, and 14-d posttreatment period. All cows were randomly assigned to a sorghum silage-based diet containing no supplemental fat (control; n=10; 138±45 d in milk) or C16:0 at 4% of ration dry matter (PALM; 98% C16:0; n=10; 136±44 d in milk). Blood and milk were collected at routine intervals. Liver and skeletal muscle tissue were biopsied at d 47 of treatment. Intravenous glucose tolerance tests (300mg/kg of body weight) were performed at d -1, 24, and 49 relative to start of treatment. The plasma and tissue concentrations of ceramide and glycosylated ceramide were determined using liquid chromatography coupled with tandem mass spectrometry. Data were analyzed as repeated measures using a mixed model with fixed effects of treatment and time, and milk yield served as a covariate. The PALM treatment increased milk yield, -corrected milk, and milk fat yield. The most abundant plasma and tissue sphingolipids detected were C24:0-ceramide, C24:0-monohexosylceramide (GlcCer), and C16:0-lactosylceramide. Plasma concentrations of total ceramide and GlcCer decreased as lactation advanced, and ceramide and GlcCer were elevated in cows fed PALM. feeding increased hepatic ceramide levels, a response not observed in skeletal muscle tissue. Plasma ceramides (e.g., C24:0-ceramide) were positively correlated with plasma NEFA and milk yield, and positively correlated with NEFA levels following a glucose challenge. Our data demonstrate a remodeled plasma and hepatic sphingolipidome in mid-lactation dairy cows fed PALM. The potential involvement in ceramide in homeorhetic nutrient partitioning to support lactation requires further consideration.Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Scale-up cultivation enhanced arachidonic accumulation by red microalgae Porphyridium purpureum.

The present study attempts to cultivate Porphyridium purpureum under different scale-up conditions for further development and commercialization of microalgae-derived PUFAs such as ARA and EPA. Different temperatures (25, 30, and 35\xa0°C) and light intensities (70, 165, and 280\xa0μmol/ms) were applied to the 50 L pilot-scale cultivation of P. purpureum in ASW. The cultivation under the light intensity of 280\xa0μmol/ms at 35\xa0°C obtained biomass concentration up to 9.52\xa0g/L, total fatty content to 56.82\xa0mg/g, and ARA content to 22.29\xa0mg/g. While the maximum EPA content of 7.00\xa0mg/g was achieved under the light intensity of 280\xa0μmol/ms at 25\xa0°C and the highest ratio of UFAs to TFAs of 74.66% was also obtained in this trial. Both biomass concentration and TFAs content were improved by increasing light intensity and temperature. Moreover, the ratio of ARA to EPA was enhanced by increasing cultivation temperature under the light intensity of 280\xa0μmol/ms. In contrast with flask culture, the conversion\xa0of linoleic (C18:2) to ARA was enhanced in scale-up culture, leading to more ARA content. Phosphate limitation enhanced the synthesis of lipid and LPUFAs. Moreover, the biomass concentration and biosynthesis of were preferred by sufficient C (NaHCO).

Keyword: energy

Improvement of extracellular lipase production by a newly isolated Yarrowia lipolytica mutant and its application in the biosynthesis of L-ascorbyl palmitate.

Yarrowia lipolytica Wt-11 producing an extracellular lipase was isolated and identified. To improve the lipase production, Y. lipolytica Wt-11 was subjected to low- ion implantation mutation breeding, and a best mutant, Y. lipolytica Mut-96, was obtained after screening. Under the optimal cultivation conditions, the scaled-up production of lipases were performed, and the lipase activity of Y. lipolytica Mut-96 was enhanced nearly 5.5-fold compared with that of Y. lipolytica Wt-11. After fermentation, the lipases were purified, and the characteristics of the purified lipases were studied. The optimum temperatures and pHs for lipases from Wt-11and Mut-96 were 30°C and 8.0, respectively. The purified lipases were stable between pH 7.0 and 8.5 and unstable at temperatures above 40°C. The lipase activities were enhanced by Ca, Ba, Mn, Fe and SDS. The synthesis of L-ascorbyl palmitate via esterification with L-ascorbic and by immobilized lipases from Wt-11 and Mut-96 in organic media was investigated, and the L-ascorbyl palmitate can be respectively produced at levels of 14.8 and 27.5g/L.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

Interfacial curvature effects on the monolayer morphology and dynamics of a clinical lung surfactant.

The morphology of surfactant monolayers is typically studied on the planar surface of a Langmuir trough, even though most physiological interfaces are curved at the micrometer scale. Here, we show that, as the radius of a clinical lung surfactant monolayer-covered bubble decreases to ∼100 µm, the monolayer morphology changes from dispersed circular liquid-condensed (LC) domains in a continuous liquid-expanded (LE) matrix to a continuous LC linear mesh separating discontinuous LE domains. The curvature-associated morphological transition cannot be readily explained by current liquid crystal theories based on isotropic domains. It is likely due to the anisotropic bending of the LC phase of the saturated phospholipids that are common to all natural and clinical lung surfactants. This continuous LC linear mesh morphology is also present on bilayer vesicles in solution. Surfactant adsorption and the dilatational modulus are also strongly influenced by the changes in morphology induced by interfacial curvature. The changes in morphology and dynamics may have physiological consequences for lung stability and function as the morphological transition occurs at alveolar dimensions.

Keyword: energy

Alterations to the microbiota-colon-brain axis in high-fat-diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated-fat diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated-fat diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy metabolism and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: energy

Impact of Brain Fatty Signaling on Peripheral Insulin Action in Mice.

Glucose homeostasis and balance are under control by peripheral and brain processes. Especially insulin signaling in the brain seems to impact whole body glucose homeostasis and interacts with fatty signaling. In humans circulating saturated fatty acids are negatively associated with brain insulin action while animal studies suggest both positive and negative interactions of fatty acids and insulin brain action. This apparent discrepancy might reflect a difference between acute and chronic fatty signaling. To address this question we investigated the acute effect of an intracerebroventricular administration on peripheral glucose homeostasis. We developed and implemented a method for simultaneous monitoring of brain activity and peripheral insulin action in freely moving mice by combining radiotelemetry electrocorticography (ECoG) and euglycemic-hyperinsulinemic clamps. This method allowed gaining insight in the early kinetics of brain fatty signaling and its contemporaneous effect on liver function , which, to our knowledge, has not been assessed so far in mice.Insulin-induced brain activity in the theta and beta band was decreased by acute intracerebroventricular application of . Peripherally it amplified insulin action as demonstrated by a significant inhibition of endogenous glucose production and increased glucose infusion rate. Moreover, our results further revealed that the brain effect of peripheral insulin is modulated by load in the brain.These findings suggest that insulin action is amplified in the periphery and attenuated in the brain by acute application. Thus, our results indicate that acute signaling in the brain may be different from chronic effects.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: energy

Energetic and conformational features linked to the monomeric and dimeric states of bovine BLG.

Bovine β-lactoglobulin (BLG) belong to the lipocalin family. This is a group of proteins involved in the binding and transporting of hydrophobic molecules. Experimental and theoretical reports have stated its complex structural behavior in solution, with coupled effects between homodimerization and ligand recognition. Nonetheless, structural evidence at the atomic level about the cause of this coupled effect has not been reported to date. To address this issue microsecond molecular dynamics (MD) simulations were combined with the molecular mechanics generalized Born surface area (MM/GBSA) approach, clustering analysis and principal component analysis (PCA), to explore the conformational complexity of BLG protein-protein self-association and (PLM) or dodecyl sulfate (SDS) ligand recognition in the monomeric and dimeric state. MD simulations, coupled to the MM/GBSA method, revealed that dimerization exerts contrasting effects on the ligand-binding capacity of BLG. Protein dimerization decreases PLM affinity, promoting dimer association. For SDS the dimeric state increases affinity, enhancing dimer dissociation. MD simulations based on PCA revealed that while few differences in the conformational subspace are observed between the free and bound monomer and dimer coupling for PLM, substantial changes are observed between the free and bound monomer and dimer coupling for SDS.Copyright © 2016 Elsevier B.V. All rights reserved.

Keyword: energy

Exploring the interactions between serum free fatty acids and fecal microbiota in obesity through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in obesity. The gut microbiota is involved in the host metabolism through the regulation of body fat storage, and a link between diet, FFA and the intestinal microbiota seems to exist. Our aim was to explore the interaction among serum FFA levels, gut microbiota, diet and obesity through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. Microbiota was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Protocatechuic Ameliorated -Induced Oxidative Damage in Endothelial Cells through Activating Endogenous Antioxidant Enzymes via an Adenosine-Monophosphate-Activated-Protein-Kinase-Dependent Pathway.

Protocatechuic (PCA, 3,4-dihydroxybenzoic ), the main metabolite of anthocyanins, is widely distributed in fruits and vegetables and has been reported to possess a strong antioxidant activity. Herein, we aimed to investigate the protective effect of PCA against high (PA)-induced oxidative damage and the underling molecular mechanisms in human umbilical vein endothelial cells (HUVECs). PCA reduced the levels of intracellular reactive oxygen species and malondialdehyde and increased the activities of endogenous antioxidant enzymes, including superoxide dismutase, glutathione peroxidase 1, and heme oxygenase 1 (HO-1). Metabolomic analysis showed that PCA affected numerous metabolites, especially some of which were related with metabolism. PCA also upregulated the phosphorylation of adenosine-monophosphate-activated protein kinase (AMPK) at Thr through activating liver kinase B1 and then promoted the expression of p-Nrf2 and HO-1. Moreover, PCA reversed the decreased expression of peroxisome proliferator-activated receptor γ coactivator 1α and significantly increased the mitochondrial density. Collectively, these results demonstrated that PCA attenuated PA-induced oxidative damage in HUVECs via an AMPK-dependent pathway.

Keyword: energy

[Influence of Fatty Acids on Oxygen Consumption in Isolated Cardiomyocytes of Rats with Ischemic or Diabetic Heart Disease].

one of the reasons of violation of the functional viability of the myocardium is considered to be the oxygen deprivation and lack of . The reason is the inhibitory effect of fatty acids on glucose oxidation. Recently, however, new data have been published proving the need for fatty acids and their importance in the maintenance and regulation of the functional activity of the myocardium in chronic pathology.to investigate the influence of free polyunsaturated and saturated fatty acids (FA) on the oxygen uptake of isolated cardiomyocytes in intact rats and animals with ischemic or diabetic heart disease.the executed non-randomized controlled study. It includied 3 groups of male rats of Wistar line (weight 250-300g) with 10 animals in each group. Myocardial infarction ("heart attack" group) was caused by ligation of the left coronary artery, diabetes ("diabetes" group)--by intraperitoneal injection of streptozotocin, and "control" group (intact animals). Myocardial infarction caused by ligation of the left coronary artery, and diabetes by intraperitoneal injection of streptozotocin. Isolated cardiac myocytes were obtained by the enzymatic method. Oxygen consumption was assessed polarographically at different saturation incubation medium with oxygen ([O₂] ≤ 8 mg/l and ([O₂] ≥ 16 mg/l). Arachidonic and acids were applied as fatty acids.It is established that the introduction of the incubation medium 20 µm arachidonic or fatty significantly increased the oxygen consumption of intact cardiomyocytes of rats. Both at the ischemic and at the diabetic injury to the heart the opposite result was obtained. The most pronounced decrease in oxygen consumption was indicated in the group with diabetes mellitus.The inhibitory effect of LCD on the rate of oxygen consumption may be associated with the influence of the ischemic or diabetic injury to the heart on the barrierfunction ofmitochondrial membranes of cardiomyocytes, the activity of membrane-associated enzymes and their associated processes.

Keyword: energy

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high- metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.

Keyword: energy

Proximate composition and fatty analysis of (L.) legume seed: implicates to both protein and essential fatty supplementation.

The high mortality rate in Bangladesh is related to poverty, which results in protein malnutrition, essential fatty deficiency and lacks in adequate vitamins, minerals and calorie. Exploring new food items with improved dietary nutrition factors may, therefore, help to decrease the mortality rate in the poor countries like Bangladesh. Accordingly, the present study was a proximate composition and fatty analysis of seed-a legume seed which is given no careful attention locally, though it might be a good source of valuable nutrition factors for both animals and humans. The purpose of the study was, therefore, to generate awareness that could also act as a good source of food components essential for good health. Proximate analysis revealed that the seed powder contained 8.47\xa0±\xa00.52% moisture; 3.50\xa0±\xa00.0.07% ash; 1.02\xa0±\xa00.06% total fat; 23.95\xa0±\xa00.15% total protein; 1.21\xa0±\xa00.16% total dietary fiber; 61.86\xa0±\xa00.70% total carbohydrate and 352.4\xa0±\xa02.66\xa0kcal/100\xa0g . Phytic content (%) was 1.014\xa0±\xa00.048. Major fatty composition (%): the essential fatty linoleic (C18:2, ω-6) was 9.50\xa0±\xa00.68, while the linolenic (C18:3, ω-3) was 1.95\xa0±\xa00.18. (C16:0), stearic (C18:0) and oleic (C18:1) were, respectively, 2.96\xa0±\xa00.19, 0.77\xa0±\xa00.04 and 1.10\xa0±\xa00.06. Lignoceric (C24:0) was 0.11\xa0±\xa00.007%. Monounsaturated palmitoleic (0.006\xa0±\xa00.0), docosapentaenoic (DPA, C22:5, ω-3) and nervonic (0.002\xa0±\xa00.0) were present in trace amounts. Arachidonic (AA, C20:4, ω-6), eicosapentaenoic (C20:5, ω-3), and docosahexaenoic (C22:6, ω-3) were not detected. The fatty profile, thus, suggests that essential omega-6 fatty linoleic (C18:3, ω-6) and omega-3 linolenic (C18:3, ω-3) were the major polyunsaturated fatty acids (PUFA) present in seed. In addition, the seed contained high amount of proteins. Finally, these results suggest that seed could be used as a good source of quality food components, including protein and essential fatty acids.

Keyword: energy

Metabolomic analysis of serum from rats following long-term intake of Chinese sausage.

Owing to the contamination of chemical pollutants, especially nitrosamines and their precursors, in Chinese sausage, long-term intake of Chinese sausage may have potential health effects.This study investigated the effects of long-term intake of Chinese sausage with different contaminations of -nitrosodimethylamine (NDMA) on rat liver and the potential biomarkers in the serum.Serum metabolomic analysis was performed by gas chromatography-mass spectrometry at weeks 7, 17, 25, and 33; simultaneously, liver histopathological examination was conducted and its relationship with the serum metabolomics was also investigated.In the study, long-term intake of Chinese sausage with different NDMA contents induced significant changes in serum metabolites and liver histopathology in rats. Metabonomic analysis showed that seven metabolites - β-alanine, 3-aminoisobutyric , aminooxyacetic , D-alanyl-D-alanine, pelargonic , (PA), and linoleic (LA) - in three sausage diet groups were significantly decreased at four time points, where three other metabolites were notably increased, which included putrescine, ethanolamine phosphate, and taurine. Among the various treatments, the NDMA (sausage-free) group demonstrated the most remarkable changes. Phenylalanine was decreased followed by an increase, and tyrosine persistently declined, both of which were elevated in the NDMA group. In addition, the histopathological result was consistent with that of the serum metabolomic analysis, and the changes in serum metabolites in each sausage diet group and the NDMA group were consistently associated with disorders of lipids, amino , and metabolism.This work indicates that excessive NDMA content in sausage may cause liver damage.

Keyword: energy

Thermo-chemo-sonic pre-digestion of waste activated sludge for yeast cultivation to extract lipids for biodiesel production.

The low cost biosynthesis of microbial lipids are an efficient feedstock to replace plant based oil for biodiesel production. The present study objective is to explore the effect of thermo-chemo-sonic pre-digestion of municipal Waste Activated Sludge (WAS) to cultivate oleaginous L.\xa0starkeyi MTCC-1400 as a model organism to produce high yield biomass and lipid. Higher Suspended Solids (SS) reduction (20 and 15.71%) and Chemical Oxygen Demand (COD) solubilization (27.6 and 22.3%) were achieved at a Specific (SE) input of 5569\xa0kJ/kg for WAS digested with NaOH and KOH, respectively. The maximum biomass of 17.52\xa0g\xa0L and lipid 64.3% dwt were attained in NaOH pre-digested sample. The analyzed lipid profile exhibited high content of (45.6%) and oleic (38.7%) which are more suitable for biofuel production. Thus, these results strongly motivate the use of pre-digested WAS as an efficient and economical substrate for biodiesel production.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Valorization of spent coffee grounds recycling as a potential alternative fuel resource in Turkey: An experimental study.

In this study, recycling of spent coffee grounds (SCG) as a potential feedstock for alternative fuel production and compounds of added value in Turkey was assessed. The average oil content was found (≈ 13% w/w). All samples (before and after extraction) were tested for scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), calorific value, surface analysis and porosity, Fourier transform infrared (FT-IR), and elemental analysis to assess their potential towards fuel properties. Elemental analysis indicated that carbon represents the highest percentages (49.59% and 46.42%, respectively), followed by nitrogen (16.7% and 15.5%), hydrogen (6.74% and 6.04%), and sulfur (0.851% and 0.561%). These results indicate that SCG can be utilized as compost, as it is rich in nitrogen. Properties of the extracted oil were examined, followed by biodiesel production. The quality of biodiesel was compared with American Society for Testing and Materials (ASTM) D6751 standards, and all the properties complied with standard specifications. The fatty compositions were analyzed by gas chromatography. It was observed that coffee waste methyl ester (CWME) is mainly composed of (35.8%) and arachidic (44.6%) acids, which are saturated fatty acids. The low degree of unsaturation provides an excellent oxidation stability (10.4\xa0hr). CWME has also excellent cetane number, higher heating value, and iodine value with poor cold flow properties. The studies also investigated blending of biodiesel with Euro diesel and butanol. Following this, a remarkable improvement in cloud and pour points of biodiesel was obtained. Spent coffee grounds after oil extraction is an ideal material for garden fertilizer, feedstock for ethanol, biogas production, and as fuel pellets. The outcome of such research work produces valuable insights on the recycling importance of SCG in Turkey.Coffee is a huge industry, and coffee has been widely used due to its refreshing properties. This industry generates large quantities of waste. Therefore, recycling of spent coffee grounds for producing alternative fuels and compounds of added value is crucial. Elemental analysis indicated that coffee waste can be utilized as compost, as it is rich in nitrogen. Coffee waste after oil extraction is an ideal feedstock for ethanol and biogas production, garden fertilizer, and as fuel pellets. The low degree of unsaturation provides excellent oxidation stability. Its biodiesel has also excellent cetane number, higher heating value, and lower iodine value.

Keyword: energy

[The physical chemical and biological features of triglycerides. The cell absorption of functionally different +oleic lipoproteins of very low and density and linoleic+linolenic lipoproteins of low density.]

The earlier insulin-independent low-density lipoproteins and more late insulin-dependent very low-density lipoproteins implement different functions at the stages of phylogenesis. The disorder of biological function of trophology, alteration of fatty acids in triglycerides, prevalence of very low-density lipoproteins over oleic very low-density lipoproteins supply mitochondria of cells with non-optimal substrate - saturated fatty for gaining , ATP synthesis. Physiologically, cells implement oleic alternative of fatty acids metabolism, oxidizing mainly ω-9 endogenous oleic mono-unsaturated fatty . The pathology of low density lipoproteins is primary deficiency of poly-unsaturated fatty acids in cells, atherosclerosis and atheromotosis of intima of arteries of elastic type with development of dense plaques from poly-unsaturated fatty acids in the form of polyethers of cholesterol. The pathology of very low-density lipoproteins includes: a) syndrome of resistance to insulin; b) pathology of phylogenetically earlier insulin-independent visceral fatty tissue - metabolic syndrome; c) pathology of phylogenetically later insulin-dependent subcutaneous adipocytes - obesity; d) secondary atherosclerosis, under cumulation of low-density lipoproteins in blood with development of atherothrombosis of intima of arteries, soft plaques rich with triglycerides. As for the prevention of disorders of transfer of fatty acids to very low-density lipoproteins and low-density lipoproteins is common in many ways - minimization of aphysiological effect of surplus amount of food, biological function of diet. The prevention at the level of population includes: a) maximal limitation of content of saturated fatty in food; b) moderate increasing of polysaturated fatty acids, ω-3 poly-saturated fatty acids predominantly; c) increasing of physical activity. The pharmaceuticals are not provided by biology in primary prevention of metabolic pandemics under aphysiological impact of environment factors.

Keyword: energy

Production of 6-O-L-Ascorbyl Palmitate by Immobilized Candida antarctica Lipase B.

L-ascorbyl palmitate (ASP) is an oil-soluble derivative of ascorbic which is used extensively in food, cosmetics industry, and medical hygiene. Enzymatic synthesis of ascorbyl palmitate in tert-butyl alcohol was carried out using indigenously immobilized lipase preparation PyCal with ascorbic and as starting material. The developed batch process under optimized reaction conditions resulted in conversion of 90% with relatively shorter reaction time of 6\xa0h. Continuous process in packed bed reactor gave conversion of 50% with space time yield of 15.46\xa0g/L/h which was found to be higher than the reported literature on enzymatic synthesis of ascorbyl palmitate. The immobilized lipase used in the present work showed good reusability. Characterization of formed ascorbyl palmitate was carried out by FTIR, MS/MS, H-NMR, and C-NMR. The enzymatic process resulted in selective synthesis of 6-O-L-ascorbyl palmitate with purity of 98.6% and no side product formation. The use of underivatized starting materials, high space time yield of 15.46\xa0g\xa0L\xa0h, high recyclability of catalyst, and no by-product formation make the overall process highly efficient and clean in terms of consumption and waste generation, respectively. The optimized reaction parameters for ascorbyl palmitate synthesis in the present study can be used as a useful reference for industrial synthesis of fatty esters of ascorbic by enzymatic route.

Keyword: energy

[Exploration about the protection mechanism of 5-hydroxy-1-methylhydantoin on paraquat poisoning model].

To investigate the effects of 5-hydroxy-1-methylhydantoin (HMH) on kidney injury induced by paraquat (PQ).Fifteen SPF healthy Kunming mice were randomly divided into normal saline (NS) control group, PQ poisoning model group and HMH intervention group, with 5 mice in each group. PQ poisoning model was challenged by one-time gavage of 30 mg/kg PQ solution. The NS group received the same amount of NS by gavage. The HMH group was given 100 mg/kg of HMH immediately after the model was made and continued to be gavaged. Mice in each group were sacrificed 1 day after HMH gavage and heart blood and renal tissue were harvested for examination. The morphological changes of renal tissue were observed under light microscope by hematoxylin-eosin (HE) staining. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in renal tissue were detected according to the instructions of the kit. The expression of heme oxygenase-1 (HO-1) and interleukin-1β (IL-1β) in renal tissues were detected by Western Blot. The serum metabolites were detected by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS), the overall distribution of each sample was observed by principal component analysis (PCA), the accuracy of the model was evaluated by multidimensional analysis orthogonal partial least squares-discriminant analysis (OPLS-DA), and the difference metabolites were screened by variable importance in the projection (VIP) value > 1.Light microscopic observation showed that: glomerular structure in NS group was clear, there was no hyperemia and inflammatory cell infiltration in renal interstitium and blood vessels. In PQ group, some glomeruli atrophy and necrosis, capillary congestion in glomeruli, infiltration of inflammatory cells around glomeruli, swelling of renal tubular epithelial cells, slight stenosis of lumen, and occasional necrosis and exfoliation of epithelial cells occurred. The degree of kidney injury in HMH group was significantly less than that in PQ group. Compared with the NS group, the content of MDA in the PQ group was significantly increased (nmol/g: 6.70±0.84 vs. 2.70±0.43, P < 0.01) and the activity of SOD was significantly decreased (kU/L: 33.30±4.66 vs. 50.20±3.23, P < 0.05), the protein expression of HO-1 and IL-1β were significantly increased (HO-1/β-actin: 1.11±0.12 vs. 0.61±0.13, IL-1β/β-actin: 0.93±0.13 vs. 0.32±0.06, both P < 0.05). Compared with the PQ group, the content of MDA in the HMH group was significantly decreased (nmol/g: 5.10±0.93 vs. 6.70±0.84, P < 0.05) and the activity of SOD was significantly increased (kU/L: 61.00±9.02 vs. 33.30±4.66, P < 0.05), the protein expression of HO-1 was significantly decreased (HO-1/β-actin: 0.77±0.07 vs. 1.11±0.12, P < 0.05), however, there was no significant difference in the protein expression of IL-1β (IL-1β/β-actin: 0.87±0.13 vs. 0.93±0.13, P > 0.05). Metabolite detection results showed that: compared with NS group, the levels of creatinine, glycine, succinic , fumaric and citric were significantly increased in the PQ group (VIP value was 1.50, 1.58, 1.64, 1.74 and 1.95 respectively, all P < 0.05), while the levels of ,α-tocopherol and 6-phosphogluconic were significantly decreased (VIP value was 1.10, 1.55 and 1.56 respectively, all P < 0.05). Compared with the PQ group, the levels of creatinine and citric were significantly decreased in the HMH group (VIP value was 1.50 and 1.86, both P < 0.05), while trans-4-hydroxy-proline, D-glyceric , 2, 6-fructose phosphate, 6-phosphate gluconic and aminomalonic were significantly increased (VIP value was 1.36, 1.55, 1.63, 1.68 and 1.76 respectively, all P < 0.05).HMH protects kidney injury caused by PQ poisoning by correcting tricarboxylic acids cycle disturbance, lipid peroxidation and energy metabolism disturbance, and its mechanism is related to the regulation of HO-1 protein expression through Nrf2 pathway.

Keyword: energy

Hepatoprotective and Antioxidant Activities of Oil from Baru Almonds ( Vog.) in a Preclinical Model of Lipotoxicity and Dyslipidemia.

The oil obtained from baru ( Vog.) almonds exhibits high value and is reported in popular medicine for the treatment of rheumatic diseases and reproductive disturbances. Although baru oil is used in domestic cuisine, the chemical characterization of this oil and its effects on lipid metabolism are still poorly understood. Therefore, this study evaluated the fatty (FA) profile and the effects of baru oil on liver and aorta in a murine model of dyslipidemia. The chromatographic profile of baru oil showed high levels of unsaturated FAs, especially oleic . Saturated FAs, such as and lignoceric acids, were found in lower amounts. Hypercholesterolemia was induced in male Wistar rats by daily administration of a lipid emulsion by gavage for 15 weeks. Biochemical and histopathological analysis were performed on serum, aorta, and liver. The results demonstrated that animals developed marked hypercholesterolemia, liver steatosis, and increased lipid peroxidation in the aorta. Treatment with baru oil attenuated lipid peroxidation and drastically reduced liver damage, especially ballooning degeneration and steatosis. By restricting vascular and hepatic injury, this oil showed potential applicability as a functional food, reinforcing its use in popular medicine and domestic cuisine.

Keyword: energy

Fatty profile of blood plasma and oviduct and uterine fluid during early and late luteal phase in the horse.

During early pregnancy, the secretome of both oviduct and uterus serves as exchange medium for signaling factors between embryo and mother and provides the embryo with nutrients. The preimplantation embryo can utilize the fatty acids (FA) therein via direct incorporation into cell membrane lipid bilayers and for production via β-oxidation. The FA concentration and composition of the oviduct (OF) and uterine fluid (UF) might be regulated by ovarian hormones to meet the changing needs of the growing embryo. In our study, we analyzed the FA profile of blood plasma (BP) and reproductive fluid samples obtained post mortem from slaughtered mares by gas chromatography mass spectrometry. Cycle stage was determined by visual evaluation of the ovary and measurement of plasma progesterone. No major effect of cycle stage on the FA profile was observed. However, the composition of FA was different between BP and both OF and UF. While linoleic, stearic, oleic and were the four most prevalent FA in both BP and reproductive fluids, the latter contained higher concentrations of arachidonic, eicosapentaenoic and dihomo-γ-linolenic . The finding suggests selective endometrial transport mechanisms from plasma into the reproductive fluids or increased endometrial synthesis of selected FA.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: energy

Optimisation, experimental validation and thermodynamic study of the sequential oil extraction and biodiesel production processes from seeds of Sterculia foetida.

Non-edible seeds are not used in any commercial applications, which implies that they can be used for biofuel applications. The present study aimed to maximise the process conditions for oil extraction and sterculia biodiesel production from Sterculia foetida (poon oil). GC-MS identified the methyl esters of sterculia oil as sterculic (32%), (15.88%), oleic (10.00%), linoleic (9.95%) and malvalic (9%). Response surface methodology (RSM) based parametric optimisation of oil extraction was carried out by choosing process variables such as sample weight, volume of solvent to seed ratio and time. The optimum sample amount of 7.5 g and the volume of solvent to seed ratio of 40 mL/g resulted in a maximum oil yield of 45.27% at 3 h. The results were statistically significant (P < 0.05) with a regression coefficient (R) of 0.9988. Furthermore, the artificial neural network (ANN) resulted in an R value greater than 0.9, which validates the RSM. Conventional optimisation of the temperature (55 °C), feedstock to methanol ratio (1:12), catalyst proportion (1.5%) and transesterification reaction time (60 min) yield 90.87% biodiesel production. The physicochemical characteristics of oil and biodiesel complied with the requirements of the ASTM standards. The rate constant and thermodynamic variables at the optimum temperature (333 K) were calculated from the experimental data. The activation (E), activation enthalpy in transition state theory (ΔH), activation entropy in transition state theory (ΔS) and Gibbs free in transition state theory (ΔG) were 37.91 kJ mol, 35.14 kJ mol, - 239.58 J mol K and 79.81 kJ mol respectively. Graphical abstract.

Keyword: energy

Increasing intake enhances milk production and prevents glucose-stimulated fatty disappearance without modifying systemic glucose tolerance in mid-lactation dairy cows.

Feeding saturated fatty acids may enhance milk yield in part by decreasing insulin sensitivity and shifting glucose utilization toward the mammary gland. Our objective was to evaluate the effects of (C16:0) on milk production and insulin sensitivity in cows. Twenty multiparous mid-lactation Holstein cows were enrolled in a study consisting of a 5-d covariate, 49-d treatment, and 14-d posttreatment period. All cows received a common sorghum silage-based diet and were randomly assigned to a diet containing no supplemental fat (control; n=10; 138±45d in milk) or C16:0 at 4% of ration DM (PALM; 98% C16:0; n=10; 136±44d in milk). Blood and milk were collected at routine intervals. Intravenous glucose tolerance tests (300mg/kg of body weight) were performed at d -1, 24, and 49 relative to start of treatment. Data were analyzed as repeated measures using a mixed model with fixed effects of treatment and time, and milk yield served as a covariate. The PALM treatment increased milk yield by wk 7. Furthermore, PALM increased milk fat yield and -corrected milk at wk 3 and 7. Changes in milk production occurred in parallel with enhanced intake. Increased milk fat yield during PALM treatment was due to increased C16:0 and C16:1 incorporation; PALM had no effect on concentration of milk components, BW, or body condition score. Two weeks posttreatment, -corrected milk and milk fat yield remained elevated in PALM-fed cows whereas yields of milk were similar between treatments. Increased milk fat yield after PALM treatment was due to increased de novo lipogenesis and uptake of preformed fatty acids. The basal concentration of nonesterified fatty acids (NEFA) in plasma increased by d 4, 6, and 8 of PALM treatment, a response not observed thereafter. Although PALM supplementation did not modify insulin, glucose, or triacylglycerol levels in plasma, total cholesterol in plasma was elevated by wk 3. Estimated insulin sensitivity was lower during the first week of PALM treatment; however, glucose disposal following glucose tolerance tests was not modified. In contrast, C16:0 feeding reduced glucose-stimulated NEFA disappearance by wk 7. Results demonstrate that increasing dietary from C16:0 for 7wk improves milk yield and milk composition without modifying systemic glucose tolerance. Reduced glucose-stimulated NEFA disappearance with C16:0 feeding and elevated circulating NEFA may reflect changes in adipose tissue insulin sensitivity.Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Consumption of individual saturated fatty acids and the risk of myocardial infarction in a UK and a Danish cohort.

The effect of individual saturated fatty acids (SFAs) on serum cholesterol levels depends on their carbon-chain length. Whether the association with myocardial infarction (MI) also differs across individual SFAs is unclear. We examined the association between consumption of individual SFAs, differing in chain lengths ranging from 4 through 18 carbons, and risk of MI.We used data from 22,050 and 53,375 participants from EPIC-Norfolk (UK) and EPIC-Denmark, respectively. Baseline SFA intakes were assessed through validated, country-specific food frequency questionnaires. Cox regression analysis was used to estimate associations between intakes of individual SFAs and MI risk, for each cohort separately.During median follow-up times of 18.8\u202fyears in EPIC-Norfolk and 13.6\u202fyears in Denmark, respectively, 1204 and 2260 MI events occurred. Mean (±SD) total SFA intake was 13.3 (±3.5) en% in EPIC-Norfolk, and 12.5 (±2.6) en% in EPIC-Denmark. After multivariable adjustment, intakes of C12:0 (lauric ) and C14:0 (myristic ) inversely associated with MI risk in EPIC-Denmark (HR upper versus lowest quintile: 0.80 (95%CI: 0.66, 0.96) for both SFAs). Intakes in the third and fourth quintiles of C4:0-C10:0 also associated with lower MI risk in EPIC-Denmark. Moreover, substitution of C16:0 () and C18:0 (stearic ) with plant proteins resulted in a reduction of MI risk in EPIC-Denmark (HR per 1 %: 0.86 (95%CI: 0.78, 0.95) and 0.87 (95%CI: 0.79, 0.96) respectively). No such associations were found in EPIC-Norfolk.The results from the present study suggest that the association between SFA and MI risk depends on the carbon chain-length of the SFA.Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: energy

Isolation and Proteomic Analysis of a Mutant with Enhanced Lipid Production by the Gamma Irradiation Method.

In this study, an enhanced lipid-producing mutant strain of the microalga was developed by gamma irradiation. To induce the mutation, was gamma irradiated at a dose of 400 Gy. After irradiation, the surviving cells were stained with Nile red. The mutant (Cr-4013) accumulating 20% more lipid than the wild type was selected. Thin-layer chromatography revealed the triglyceride and free fatty contents to be markedly increased in Cr-4013. The major fatty acids identified were , oleic , linoleic , and linolenic . Random amplified polymeric DNA analysis showed partial genetic modifications in Cr-4013. To ascertain the changes of protein expression in the mutant strain, two-dimensional electrophoresis was conducted. These results showed that gamma radiation could be used for the development of efficient microalgal strains for lipid production.

Keyword: energy

Exploring Strategies for Labeling Viruses with Gold Nanoclusters through Non-equilibrium Molecular Dynamics Simulations.

Biocompatible gold nanoclusters can be utilized as contrast agents in virus imaging. The labeling of viruses can be achieved noncovalently but site-specifically by linking the cluster to the hydrophobic pocket of a virus via a lipid-like pocket factor. We have estimated the binding affinities of three different pocket factors of echovirus 1 (EV1) in molecular dynamics simulations combined with non-equilibrium free- calculations. We have also studied the effects on binding affinities with a pocket factor linked to the AupMBA nanocluster in different protonation states. Although the absolute binding affinities are over-estimated for all the systems, the trend is in agreement with recent experiments.3 Our results suggest that the natural pocket factor () can be replaced by molecules pleconaril (drug) and its derivative Kirtan1 that have higher estimated binding affinities. Our results also suggest that including the gold nanocluster does not decrease the affinity of the pocket factor to the virus, but the affinity is sensitive to the protonation state of the nanocluster, i.e., to pH conditions. The methodology introduced in this work helps in the design of optimal strategies for gold-virus bioconjugation for virus detection and manipulation.

Keyword: energy

: Physiological Role, Metabolism and Nutritional Implications.

(PA) has been for long time negatively depicted for its putative detrimental health effects, shadowing its multiple crucial physiological activities. PA is the most common saturated fatty accounting for 20-30% of total fatty acids in the human body and can be provided in the diet or synthesized endogenously via lipogenesis (DNL). PA tissue content seems to be controlled around a well-defined concentration, and changes in its intake do not influence significantly its tissue concentration because the exogenous source is counterbalanced by PA endogenous biosynthesis. Particular physiopathological conditions and nutritional factors may strongly induce DNL, resulting in increased tissue content of PA and disrupted homeostatic control of its tissue concentration. The tight homeostatic control of PA tissue concentration is likely related to its fundamental physiological role to guarantee membrane physical properties but also to consent protein palmitoylation, palmitoylethanolamide (PEA) biosynthesis, and in the lung an efficient surfactant activity. In order to maintain membrane phospholipids (PL) balance may be crucial an optimal intake of PA in a certain ratio with unsaturated fatty acids, especially PUFAs of both n-6 and n-3 families. However, in presence of other factors such as positive balance, excessive intake of carbohydrates (in particular mono and disaccharides), and a sedentary lifestyle, the mechanisms to maintain a steady state of PA concentration may be disrupted leading to an over accumulation of tissue PA resulting in dyslipidemia, hyperglycemia, increased ectopic fat accumulation and increased inflammatory tone via toll-like receptor 4. It is therefore likely that the controversial data on the association of dietary PA with detrimental health effects, may be related to an excessive imbalance of dietary PA/PUFA ratio which, in certain physiopathological conditions, and in presence of an enhanced DNL, may further accelerate these deleterious effects.

Keyword: energy

Resistance exercise training and in\xa0vitro skeletal muscle oxidative capacity in older adults.

Whether resistance exercise training (RET) improves skeletal muscle substrate oxidative capacity and reduces mitochondrial production of reactive oxygen species in older adults remains unclear. To address this, 19 older males (≥60\xa0years) were randomized to a RET (n\xa0=\xa011) or to a waitlist control group (n\xa0=\xa08) that remained sedentary for 12\xa0weeks. RET was comprised of three upper body and four lower body movements on resistance machines. One set of 8-12 repetitions to failure of each movement was performed on three nonconsecutive days/week. Improvements in chest press and leg press strength were assessed using a three-repetition maximum (3 RM). Body composition was assessed via dual X-ray absorptiometry. Muscle biopsies were obtained from the vastus lateralis muscle at baseline and at both 3\xa0weeks and 12\xa0weeks. Palmitate and pyruvate oxidation rates were measured from the (14)CO2 produced from [1-(14)C] and [U-(14)C] pyruvate, respectively, during incubation of muscle homogenates. PGC-1α, TFAM, and PPARδ levels were quantified using qRT-PCR Citrate synthase (CS) and β-HAD activities were determined spectrophotometrically. Mitochondrial production of reactive oxygen species (ROS) were assessed using the Amplex Red Hydrogen Peroxide/Peroxidase assay. There were no significant changes in body weight or body composition following the intervention. Chest press and leg press strength (3RM) increased ~34% (both P\xa0<\xa00.01) with RET There were no significant changes in pyruvate or fatty oxidation or in the expression of target genes with the intervention. There was a modest increase (P\xa0<\xa00.05) in βHAD activity with RET at 12\xa0weeks but the change in CS enzyme activity was not significant. In addition, there were no significant changes in ROS production in either group following RET Taken together, the findings of this study suggest that 12\xa0weeks of low volume RET does not increase skeletal muscle oxidative capacity or reduce ROS production in older adults.© 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

Keyword: energy

Dietary modulation of homoeostasis and metabolic-inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-inflammation, within the context of obesity, type-2 diabetes (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular metabolism is a key determinant of inflammation. Whilst metabolic-inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: energy

Variant Amino Residues Alter the Enzyme Activity of Peanut Type 2 Diacylglycerol Acyltransferases.

Diacylglycerol acyltransferase (DGAT) catalyzes the final step in triacylglycerol (TAG) biosynthesis via the acyl-CoA-dependent acylation of diacylglycerol. This reaction is a major control point in the Kennedy pathway for biosynthesis of TAG, which is the most important form of stored metabolic in most oil-producing plants. In this study, Arachis hypogaea type 2 DGAT (AhDGAT2) genes were cloned from the peanut cultivar \'Luhua 14.\' Sequence analysis of 11 different peanut cultivars revealed a gene family of 8 peanut DGAT2 genes (designated AhDGAT2a-h). Sequence alignments revealed 21 nucleotide differences between the eight ORFs, but only six differences result in changes to the predicted amino (AA) sequences. A representative full-length cDNA clone (AhDGAT2a) was characterized in detail. The biochemical effects of altering the AhDGAT2a sequence to include single variable AA residues were tested by mutagenesis and functional complementation assays in transgenic yeast systems. All six mutant variants retained enzyme activity and produced lipid droplets in vivo. The N6D and A26P mutants also displayed increased enzyme activity and/or total cellular fatty (FA) content. N6D mutant mainly increased the content of palmitoleic , and A26P mutant mainly increased the content of . The A26P mutant grew well both in the presence of oleic and C18:2, but the other mutants grew better in the presence of C18:2. AhDGAT2 is expressed in all peanut organs analyzed, with high transcript levels in leaves and flowers. These levels are comparable to that found in immature seeds, where DGAT2 expression is most abundant in other plants. Over-expression of AhDGAT2a in tobacco substantially increased the FA content of transformed tobacco seeds. Expression of AhDGAT2a also altered transcription levels of endogenous tobacco lipid metabolic genes in transgenic tobacco, apparently creating a larger carbon \'sink\' that supports increased FA levels.

Keyword: energy

Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in lipid metabolism. We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, oxygen consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high-fat diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased fat mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less oxygen, indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo lipogenesis and results in dyslipidaemia and obesity.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: energy

Prepartum fatty supplementation in sheep I. Eicosapentaenoic and docosahexaenoic supplementation do not modify ewe and lamb metabolic status and performance through weaning.

Fatty acids are involved in the regulation of many physiological pathways, including those involved in gene expression and metabolism. Through effects on these pathways, fatty acids may have lifelong impacts on offspring development and metabolism via maternal supplementation. Therefore, our objective was to investigate the impact of supplementing a source of omega-3 fatty acids eicosapentaenoic (EPA) and docosahexaenoic (DHA) during late gestation on productive and metabolic responses of ewes and their offspring. Eighty-four gestating ewes (28 pens) were blocked and randomly assigned to a diet with 0.39% added fat during the last 50 d of gestation (d -0). The fat sources were Ca salts of a fatty distillate (PFAD) or EPA + DHA. After lambing (d 1), all ewes and lambs were placed on the same pasture. The ewes were weighed and BCS was measured on d -50, -20, 30, and 60 (weaning) of the experiment. Blood samples were taken from the ewes on d -50, -20, 1 (lambing), 30, and 60. Milk yield and composition were measured at 30 d postpartum. Lambs were weighed and bled at d 1, 30, and 60, and ADG was calculated. All plasma samples were analyzed for glucose and NEFA. Ghrelin, prostaglandin E metabolites (PGEM), and the prostaglandin D2 metabolite 11β-PGF2α were measured in d -20 ewe samples. Insulin and adropin were measured in lamb samples at d 60. There was no difference on ewe BW (P = 0.48) or BCS (P = 0.55), or plasma concentrations of glucose (P = 0.57), NEFA (P = 0.44), ghrelin (P = 0.36), PGEM (P = 0.32), and 11β-PGF2α (P = 0.86) between ewes supplemented with PFAD or EPA + DHA. Neither milk yield nor its composition was different (P > 0.10) among treatments. Lambs born from ewes supplemented with PFAD or EPA + DHA did not have different BW (P = 0.22), ADG (P = 0.21) or plasma NEFA (P = 0.52), glucose (P = 0.50), insulin (P = 0.59), and adropin (P = 0.72) concentrations. These results suggest that supplementation of EPA and DHA during late gestation did not affect ewe metabolic profile or milk production. Lamb performance and metabolism through weaning were not affected by maternal supplementation with an enriched source of EPA and DHA.

Keyword: energy

[THE EXCESS OF FATTY IN FOOD AS MAIN CAUSE OF LIPOIDOSIS OF INSULIN-DEPENDENT CELLS: SKELETAL MYOCYTES, CARDIO-MYOCYTES, PERIPORTAL HEPATOCYTES, KUPFFER MACROPHAGES AND B-CELLS OF PANCREAS].

In phylogenesis, becoming of biologicalfunctions and biological reactions proceeds with the purpose ofpermanent increasing of "kinetic perfection ". The main role belongs to factors ofphysical, chemical and biological kinetics, their evaluation using systemic approach technique under permanent effect of natural selection. The late-in-phylogenesis insulin, proceeded with, in development of biological function of locomotion, specialization of insulin-dependent cells: skeletal myocytes, syncytium of cardiomyocytes, subcutaneous adipocytes, periportal hepatocytes, Kupffer\'s macrophages and β-cells of islets of pancreas. The insulin initiated formation of new, late in phylogenesis, large pool of fatty cells-subcutaneous adipocytes that increased kinetic parameters of biological function of locomotion. In realization of biological function of locomotion only adipocytes absorb exogenous mono unsaturated and saturated fatty acids in the form of triglycerides in composition of oleic and lipoproteins of very low density using apoE/B-100 endocytosis. The rest of insulin-dependent cells absorb fatty acids in the form of unesterified fatty acids from associates with albumin and under effect of CD36 of translocase offatty acids. The insulin in all insulin-depended cells inhibits biological reaction of lipolysis enhancing contributing into development of lipoidosis. The insulin expresses transfer offatty acids in the form of unsaturated fatty acids from adipocytes into matrix of mitochondria. The insulin supplies insulin-dependent cells with substrates for acquiring subject to that in pool of unsaturated fatty acids in adipocytes prevails hydrophobic unsaturated fatiy that slowly passes into matrix through external membrane ofmitochondria; oxidases of mitochondria so slowly implement its β-oxidation that content of exogenous unsaturatedfatty can\'t be higher than phylogenetic, physiological level - 15% of all amount offatty acids transferring to insulin-dependent cells. The insulin can\'t both to decrease content of exogenous fatty and inhibit lipolysis in visceral fatty cells of omentum.

Keyword: energy

GC-MS metabolomics reveals disturbed metabolic pathways in primary mouse hepatocytes exposed to subtoxic levels of 3,4-methylenedioxymethamphetamine (MDMA).

3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is a well-known hepatotoxic drug. Although its toxicity has been thoroughly studied at high concentrations, there is still insufficient knowledge on possible alterations of cell function at subtoxic concentrations, which are in fact more representative concentrations of intoxication scenarios. In this study, a gas chromatography-mass spectrometry (GC-MS) metabolomics approach was used to investigate the metabolic changes in primary mouse hepatocytes (PMH) exposed to two subtoxic concentrations of MDMA (LC and LC) for 24\xa0h. Metabolomic profiling of both intracellular metabolites and volatile metabolites in the extracellular medium of PMH was performed. Multivariate analysis showed that the metabolic pattern of cells exposed to MDMA discriminates from the controls in a concentration-dependent manner. Exposure to LC MDMA induces a significant increase in some intracellular metabolites, including oleic and , and a decrease in glutamate, aspartate, 5-oxoproline, fumarate, malate, phosphoric , α-ketoglutarate and citrate. Extracellular metabolites such as acetophenone, formaldehyde, pivalic , glyoxal and 2-butanone were found significantly increased after exposure to MDMA, compared to controls, whereas 4-methylheptane, 2,4-dimethyl-1-heptene, nonanal, among others, were found significantly decreased. The panel of discriminatory metabolites is mainly involved in tricarboxylic (TCA) cycle, fatty metabolism, glutamate metabolism, antioxidant defenses and possibly changes in the liver enzyme machinery. Overall, these results highlight the potential of the intra- and extracellular metabolome to study alterations triggered by subtoxic concentrations of MDMA in hepatic cell functions, which represents a more realistic appraisal of early toxicity events posed by exposure to this drug. In addition, these results also revealed some metabolites that may be used as potential biomarkers indicative of early events in the hepatotoxicity induced by MDMA.

Keyword: energy

P66Shc-Induced MicroRNA-34a Causes Diabetic Endothelial Dysfunction by Downregulating Sirtuin1.

Diabetes mellitus causes vascular endothelial dysfunction and alters vascular microRNA expression. We investigated whether endothelial microRNA-34a (miR-34a) leads to diabetic vascular dysfunction by targeting endothelial sirtuin1 (Sirt1) and asked whether the oxidative stress protein p66Shc governs miR-34a expression in the diabetic endothelium.MiR-34a is upregulated, and Sirt1 downregulated, in aortic endothelium of db/db and streptozotocin-induced diabetic mice. Systemic administration of miR-34a inhibitor, or endothelium-specific knockout of miR-34a, prevents downregulation of aortic Sirt1 and rescues impaired endothelium-dependent aortic vasorelaxation induced by diabetes mellitus. Moreover, overexpression of Sirt1 mitigates impaired endothelium-dependent vasorelaxation caused by miR-34a mimic ex vivo. Systemic infusion of miR-34a inhibitor or genetic ablation of endothelial miR-34a prevents downregulation of endothelial Sirt1 by high glucose. MiR-34a is upregulated, Sirt1 is downregulated, and oxidative stress (hydrogen peroxide) is induced in endothelial cells incubated with high glucose or the free fatty palmitate in vitro. Increase of hydrogen peroxide and induction of endothelial miR-34a by high glucose or palmitate in vitro is suppressed by knockdown of p66shc. In addition, overexpression of wild-type but not redox-deficient p66Shc upregulates miR-34a in endothelial cells. P66Shc-stimulated upregulation of endothelial miR-34a is suppressed by cell-permeable antioxidants. Finally, mice with global knockdown of p66Shc are protected from diabetes mellitus-induced upregulation of miR-34a and downregulation of Sirt1 in the endothelium.These data show that hyperglycemia and elevated free fatty acids in the diabetic milieu recruit p66Shc to upregulate endothelial miR-34a via an oxidant-sensitive mechanism, which leads to endothelial dysfunction by targeting Sirt1.© 2016 American Heart Association, Inc.

Keyword: energy

Altering the ratio of dietary , stearic, and oleic acids in diets with or without whole cottonseed affects nutrient digestibility, partitioning, and production responses of dairy cows.

The objective of this study was to evaluate the effects of varying the ratio of dietary (C16:0), stearic (C18:0), and oleic (cis-9 C18:1) acids in basal diets containing soyhulls or whole cottonseed on nutrient digestibility, partitioning, and production response of lactating dairy cows. Twenty-four mid-lactation multiparous Holstein cows were used in a split-plot Latin square design. Cows were allocated to a main plot receiving either a basal diet with soyhulls (SH, n = 12) or a basal diet with whole cottonseed (CS, n = 12) that was fed throughout the experiment. Within each plot a 4 × 4 Latin square arrangement of treatments was used in 4 consecutive 21-d periods. Treatments were (1) control (CON; no supplemental fat), (2) high C16:0 supplement [PA; fatty (FA) supplement blend provided ∼80% C16:0], (3) C16:0 and C18:0 supplement (PA+SA; FA supplement blend provided ∼40% C16:0 + ∼40% C18:0), and (4) C16:0 and cis-9 C18:1 supplement (PA+OA; FA supplement blend provided ∼45% C16:0 + ∼35% cis-9 C18:1). Interactions between basal diets and FA treatments were observed for dry matter intake (DMI) and milk yield. Among the SH diets, PA and PA+SA increased DMI compared with CON and PA+OA treatments, whereas in the CS diets PA+OA decreased DMI compared with CON. The PA, PA+SA, and PA+OA treatments increased milk yield compared with CON in the SH diets. The CS diets increased milk fat yield compared with the SH diets due to the greater yield of de novo and preformed milk FA. The PA treatment increased milk fat yield compared with CON, PA+SA, and PA+OA due to the greater yield of mixed-source (16-carbon) milk FA. The PA treatment increased 3.5% fat-corrected milk compared with CON and tended to increase it compared with PA+SA and PA+OA. The CS diets increased body weight (BW) change compared with the SH diets. Additionally, PA+OA tended to increase BW change compared with CON and PA and increased it in comparison with PA+SA. The PA and PA+OA treatments increased dry matter and neutral detergent fiber digestibility compared with PA+SA and tended to increase them compared with CON. The PA+SA treatment reduced 16-carbon, 18-carbon, and total FA digestibility compared with the other treatments. The CS diets increased partitioning toward body reserves compared with the SH diets. The PA treatment increased partitioning toward milk compared with CON and PA+OA and tended to increase it compared with PA+SA. In contrast, PA+OA increased partitioned to body reserves compared with PA and PA+SA and tended to increase it compared with CON. In conclusion, milk yield responses to different combinations of FA were affected by the addition of whole cottonseed in the diet. Among the combinations of C16:0, C18:0, and cis-9 C18:1 evaluated, fat supplements with more C16:0 increased output in milk, whereas fat supplements with more cis-9 C18:1 increased storage in BW. The combination of C16:0 and C18:0 reduced nutrient digestibility, which most likely explains the lower performance observed compared with other treatments.Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Chronic Heat Stress Induces Acute Phase Responses and Serum Metabolome Changes in Finishing Pigs.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN; 22 °C) or heat stress (HS, 30 °C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values >1 and < 0.05). These metabolites are involved in carbohydrate, amino , fatty , amines metabolism, and gut microbiome-derived metabolism. Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic were decreased, indicating insufficient source of fuel for supply, resulting in negative balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty (myristic , , and linoleic ) and short-chain fatty acids (3-hydroxybutanoic and maleic ) suggested fat decomposition compensating for shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic were significantly correlated with the levels of acute phase proteins (HP, LBP, α2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.

Keyword: energy

Palmitoleic (16:1n7) increases oxygen consumption, fatty oxidation and ATP content in white adipocytes.

We have recently demonstrated that palmitoleic (16:1n7) increases lipolysis, glucose uptake and glucose utilization for production in white adipose cells. In the present study, we tested the hypothesis that palmitoleic modulates bioenergetic activity in white adipocytes.For this, 3\xa0T3-L1 pre-adipocytes were differentiated into mature adipocytes in the presence (or absence) of (16:0) or palmitoleic (16:1n7) at 100 or 200\xa0μM. The following parameters were evaluated: lipolysis, lipogenesis, fatty (FA) oxidation, ATP content, oxygen consumption, mitochondrial mass, citrate synthase activity and protein content of mitochondrial oxidative phosphorylation (OXPHOS) complexes.Treatment with 16:1n7 during 9\xa0days raised basal and isoproterenol-stimulated lipolysis, FA incorporation into triacylglycerol (TAG), FA oxidation, oxygen consumption, protein expression of subunits representing OXPHOS complex II, III, and V and intracellular ATP content. These effects were not observed in adipocytes treated with 16:0.Palmitoleic , by concerted action on lipolysis, FA esterification, mitochondrial FA oxidation, oxygen consumption and ATP content, does enhance white adipocyte expenditure and may act as local hormone.

Keyword: energy

Short-term starvation is a strategy to unravel the cellular capacity of oxidizing specific exogenous/endogenous substrates in mitochondria.

Mitochondrial oxidation of nutrients is tightly regulated in response to the cellular environment and changes in demands. studies evaluating the mitochondrial capacity of oxidizing different substrates are important for understanding metabolic shifts in physiological adaptations and pathological conditions, but may be influenced by the nutrients present in the culture medium or by the utilization of endogenous stores. One such influence is exemplified by the Crabtree effect (the glucose-mediated inhibition of mitochondrial respiration) as most experiments are performed in glucose-containing media. Here, using high-resolution respirometry, we evaluated the oxidation of endogenous or exogenous substrates by cell lines harboring different metabolic profiles. We found that a 1-h deprivation of the main energetic nutrients is an appropriate strategy to abolish interference of endogenous or undesirable exogenous substrates with the cellular capacity of oxidizing specific substrates, namely glutamine, pyruvate, glucose, or palmitate, in mitochondria. This approach primed mitochondria to immediately increase their oxygen consumption after the addition of the exogenous nutrients. All starved cells could oxidize exogenous glutamine, whereas the capacity for oxidizing palmitate was limited to human hepatocarcinoma Huh7 cells and to C2C12 mouse myoblasts that differentiated into myotubes. In the presence of exogenous glucose, starvation decreased the Crabtree effect in Huh7 and C2C12 cells and abrogated it in mouse neuroblastoma N2A cells. Interestingly, the fact that the Crabtree effect was observed only for mitochondrial basal respiration but not for the maximum respiratory capacity suggests it is not caused by a direct effect on the electron transport system.© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: energy

Increased FNDC5/Irisin expression in human hepatocellular carcinoma.

The fibronectin type III domain containing 5 (FNDC5)/Irisin, a novel -regulating hormone, is associated with lipid and carbohydrate metabolism. It is produced in low amounts by normal hepatic tissue, while in human hepatocellular carcinoma (HCC), in which aberrant de novo lipogenesis (DNL) occurs, the hepatic expression of FNDC5/Irisin is still unknown. The gene expression of FNDC5/Irisin, associated to key regulators of DNL, inflammation and cancer progression was evaluated in liver tissue of 18 patients with HCC undergoing liver transplantation and of 18 deceased donors. Hepatic mRNA expression of FNDC5/Irisin and stearoyl-CoA desaturase (SCD-1), main enzymatic regulator of DNL, were significantly higher in HCC patients than in donors (p<0.0001 and p=0.015, respectively). The hepatic mRNA expression of the neurogenic locus notch homolog protein 1 (NOTCH1) tended to be higher in HCC patients than in donors (p=0.06). Only in HCC patients, hepatic FNDC5/Irisin strongly correlated with the transcription factor sterol regulatory element-binding factor 1, SCD-1, NOTCH1, tumor necrosis factor-α and Interleukin-6 mRNA expression. Further, in HCC patients, FNDC5/Irisin mRNA tended to correlate to plasma lipid profile namely triglycerides, /linoleic and polyunsaturated fatty /saturated fatty ratios. In conclusion, HCC-liver tissue over-expressed FNDC5/Irisin in association with gene expression of mediators involved in lipogenesis, inflammation and cancer, suggesting a possible protective role of the hormone from the liver damage.Copyright © 2016 Elsevier Inc. All rights reserved.

Keyword: energy

Environmental alterations in biofuel generating molecules in Zilla spinosa.

Now days, production of fuels and petrochemicals from renewable lignocellulosic biomass is an indispensable issue to meet the growing demand. Meanwhile, the changes in the climate and soil topography influence the growth and development as well as canopy level of the lignocellulosic biomass. In this study, Zilla spinosa Turr (Zilla) plants with similar age and size were collected from three main sectors (upstream, midstream, and downstream) of Wadi Hagul during spring (April) and summer (July) seasons. Environmental stresses evoked reduction in the trapping pigments concomitant with increments in chlorophyll fluorescence in summer harvested plants particularly at downstream. Furthermore, the biofuels generating compounds including carbohydrate, lignin, and lipid making the plant biomasses are greatly affected by environmental conditions. Greater amount of lignin was estimated in summer harvested Z. spinosa shoots particularly at downstream. Moreover, the total oil content which is a promising source of biodiesel was considerably decreased during summer season particularly at downstream. The physical properties of the lipids major constituent fatty methyl esters determine the biofuel properties and contribute in the adaptation of plants against environmental stresses. Hence, the analysis of fatty profile showed significant modifications under combined drought and heat stress displayed in the summer season. The maximum increase in saturated fatty levels including tridecanoic (C13:0), pentadeanoic (C15:0), (C16:0), and stearic (C18:0) were estimated in spring harvested Z. spinosa aerial portions particularly at midstream. In spite of the reduction in the total oil content, a marked increase in the value of unsaturated to saturated fatty acids ratio and thereby the unsaturation index were achieved during the dry summer period. Henceforth, these seasonal and spatial variations in fatty acids profiles may contribute in the acclimatization of Z. spinosa plants to soil water scarcity associated with heat stress experienced during summer. In addition, the alterations in the fatty profiles may match biofuel requirements. In conclusion, the most adequate growing season (spring) will be decisive for achieving high lipid productivity associated with improved biofuel quality in terms of high saturated fatty acids percentage that improves its cetane number. However, the dry summer season enhanced the accumulation of greater amount of lignin that may enhance the biodiesel quantity.

Keyword: energy

Chemical composition, in vitro gas production, methane production and fatty profile of canola silage (Brassica napus) with four levels of molasses.

The objective of this study was to investigate the effect of four levels of molasses on chemical composition, in vitro digestibility, methane production and fatty profile of canola silages. A canola (Brassica napus var. Monty) crop was established in a small-scale agricultural farm and harvested 148\xa0days after sowing. Four levels of molasses were tested with respect to the fresh weight (1.5\xa0kg); these were 1% (CS-1), 2% (CS-2), 3% (CS-3) and 4% (CS-4) molasses, and 0% molasses (CS-0) was included as a control. A total of 45 microsilages were prepared using PVC pipes (4\xa0in. of diameter × 20\xa0cm of length), and the forage was compressed using a manual press. The effects of control and treatments were tested using the general linear model Y\u2009=\u2009μ\u2009+\u2009T\u2009+\u2009E. The linolenic (C18:3n3), (C16:0) and linoleic methyl ester (C18:2n6c) accounted for 30%, 21% and 10.5% of total fatty acids, respectively; the fermentation parameters and in vitro methane production were not affected (P\u2009>\u20090.05) by treatments; in vitro digestibility decreased significantly (P\u2009<\u20090.05) as the level of molasses increased. It was concluded that CS-4 improved the DM content by 9% and showed high content of linolenic methyl ester. The gross of canola silages could favour the oleic methyl ester.

Keyword: energy

High α-tocopherol dosing increases lipid metabolism by changing redox state in damaged rat gastric mucosa and liver after ethanol treatment.

Regeneration of ethanol-injured rat gastric mucosa must undergo changes in major metabolic pathways to achieve DNA replication and cell proliferation. These events are highly dependent on glucose utilization and inhibited by vitamin E (VE) (α-tocopherol) administration. Therefore, the present study aimed at assessing lipid metabolism in the gastric mucosa and ethanol-induced gastric damage and the effect of α-tocopherol administration. For this, rates of fatty β-oxidation and lipogenesis were tested in gastric mucosa samples. Through histological analysis, we found loss of the mucosa\'s superficial epithelium, which became gradually normalized during the recovery period. Proliferation of gastric mucosa occurred with augmented formation of β-oxidation by-products, diminished synthesis of triacylglycerols (TGs), as well as of phospholipids, and a reduced cytoplasmic NAD/NADH ratio, whereas the mitochondrial redox NAD/NADH ratio was much less affected. In addition, α-tocopherol increased utilization in the gastric mucosa, which was accompanied by the induction of \'mirror image\' effects on the cell redox state, reflected in an inhibited cell gastric mucosa proliferation by the vitamin administration. In conclusion, the present study shows, for the first time, the role of lipid metabolism in the adaptive cell gastric mucosa changes that drive proliferation after a chronic insult. Moreover, α-tocopherol increased gastric mucosa utilization of associated with production. These events could be associated with its antioxidant properties in co-ordination with regulation of genes and cell pathways, including changes in the cell NAD/NADH redox state.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: energy

Enhanced lipid extraction from oleaginous yeast biomass using ultrasound assisted extraction: A greener and scalable process.

Soaring demand for alternative fuels has been gaining wide interest due to depletion of conventional fuel, increasing petroleum prices and greenhouse gas emissions. Biodiesel, an alternative fuel, derived from oleaginous microbes has been promising because of short incubation time and easy to scale up. Oleaginous yeast Trichosporon sp. is capable of utilizing glycerol and agro-residues for enhanced lipid synthesis. Lipid extraction from Trichosporon sp. biomass showed highest lipid content with ultrasonic assisted extraction (43\u202f±\u202f0.33%, w/w) coupled with process parameters than the conventional Soxhlet (30\u202f±\u202f0.28%, w/w) and Binary solvent [choloroform:methanol, (2:1, v/v)] methods (36\u202f±\u202f0.38%, w/w), respectively. The standardized process parameters of ultrasonic assisted extraction coupled with chloroform/methanol solvent system resulted 95-97% of conversion efficiency in 20\u202fmin at 30\u202f°C with a frequency of 50\u202fHz and 2800\u202fW power, respectively. Enzymatic transesterification of yeast biomass lipid obtained 85% of fatty methyl esters that are predominant with oleic methyl ester followed by and stearic methyl esters, respectively. These results substantiate that the ultrasonic assisted extraction is a potential green extraction technique that had reduced time, energy and solvent consumption without compromising on lipid quality. Deploying this green extraction technique could make the biodiesel production process inexpensive and eco-friendly.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Study of the Thermal Properties and the Fire Performance of Flame Retardant-Organic PCM in Bulk Form.

The implementation of organic phase change materials (PCMs) in several applications such as heating and cooling or building comfort is an important target in thermal storage (TES). However, one of the major drawbacks of organic PCMs implementation is flammability. The addition of flame retardants to PCMs or shape-stabilized PCMs is one of the approaches to address this problem and improve their final deployment in the building material sector. In this study, the most common organic PCM, Paraffin RT-21, and fatty acids mixtures of capric (CA), myristic (MA), and (PA) in bulk, were tested to improve their fire reaction. Several flame retardants, such as ammonium phosphate, melamine phosphate, hydromagnesite, magnesium hydroxide, and aluminum hydroxide, were tested. The properties of the improved PCM with flame retardants were characterized by thermogravimetric analyses (TGA), the dripping test, and differential scanning calorimetry (DSC). The results for the dripping test show that fire retardancy was considerably enhanced by the addition of hydromagnesite (50 wt %) and magnesium hydroxide (50 wt %) in fatty acids mixtures. This will help the final implementation of these enhanced PCMs in building sector. The influence of the addition of flame retardants on the melting enthalpy and temperatures of PCMs has been evaluated.

Keyword: energy

Metabolomic study for monitoring of biomarkers in mouse plasma with asthma by gas chromatography-mass spectrometry.

Asthma is a multifaceted chronic disease caused by an alteration of various genetic and environmental factors that is increasing in incidence worldwide. However, the biochemical mechanisms regarding asthma are not completely understood. Thus, we performed of metabolomic study for understanding of the biochemical events by monitoring of altered metabolism and biomarkers in asthma. In mice plasma, 27 amino acids(AAs), 24 fatty acids(FAs) and 17 organic acids(OAs) were determined by ethoxycarbonyl(EOC)/methoxime(MO)/tert-butyldimethylsilyl(TBDMS) derivatives with GC-MS. Their percentage composition normalized to the corresponding mean levels of control group. They then plotted as star symbol patterns for visual monitoring of altered metabolism, which were characteristic and readily distinguishable in control and asthma groups. The Mann-Whitney test revealed 25 metabolites, including eight AAs, nine FAs and eight OAs, which were significantly different (p<0.05), and orthogonal partial least-squares-discriminant analysis revealed a clear separation of the two groups. In classification analysis, and methionine were the main metabolites for discrimination between asthma and the control followed by pipecolic, lactic, α-ketoglutaric, and linoleic acids for high classification accuracy as potential biomarkers. These explain the metabolic disturbance in asthma for AAs and FAs including intermediate OAs related to the metabolism in the TCA cycle.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: energy

Physicochemical properties, nutritional value and techno-functional properties of goldenberry (Physalis peruviana) waste powder concise title: Composition of goldenberry juice waste.

Goldenberry waste powder, contained 5.87% moisture, 15.89% protein, 13.72% fat, 3.52% ash, 16.74% dietary fiber and 61% carbohydrates. Potassium (560\u202fmg/100\u202fg) was the predominant element followed by sodium (170\u202fmg/100\u202fg) and phosphorus (130\u202fmg/100\u202fg). Amino analysis gave high levels of cystine/methionine, histidine and tyrosine/phenylalanine. Goldenberry waste powder had good levels of the techno-functional properties including water absorption index, swelling index, foaming capacity and stability (3.38\u202fg/g, 5.24\u202fml/g, 4.09 and 72.0%, respectively). Fatty acids profile showed that linoleic was the predominant fatty followed by oleic, and stearic acids. Iodine value (109.5\u202fg/100\u202fg of oil), value (2.36\u202fmg KOH/g of oil), saponification value (183.8\u202fmg KOH/g of oil), peroxide value (8.2\u202fmeq/kg of oil) and refractive index (1.4735) were comparable to those of soybean and sunflower oils. Goldenberry waste oil exhibited absorbance in the UV range at 100-400\u202fnm.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

Thermodynamic versus non-equilibrium stability of monolayers in calcium-enriched sea spray aerosol proxy systems.

Of the major cations in seawater (Na+, Mg2+, Ca2+, K+), Ca2+ is found to be the most enriched in fine sea spray aerosols (SSA). In this work, we investigate the binding of Ca2+ to the carboxylic headgroup of (PA), a marine-abundant fatty , and the impact such binding has on the stability of PA monolayers in both equilibrium and non-equilibrium systems. A range of Ca2+ conditions from 10 μM to 300 mM was utilized to represent the relative concentration of Ca2+ in high and low relative humidity aerosol environments. The CO2- stretching modes of PA detected by surface-sensitive infrared reflection-absorption spectroscopy (IRRAS) reveal ionic binding motifs of the Ca2+ ion to the carboxylate group with varying degrees of hydration. Surface tensiometry was used to determine the thermodynamic equilibrium spreading pressure (ESP) of PA on the various aqueous CaCl2 subphases. Up to concentrations of 1 mM Ca2+, each system reached equilibrium, and Ca2+:PA surface complexation gave rise to a lower state revealed by elevated surface pressures relative to water. We show that PA films are not thermodynamically stable at marine aerosol-relevant Ca2+ concentrations ([Ca2+] ≥ 10 mM). IRRAS and vibrational sum frequency generation (VSFG) spectroscopy were used to investigate the surface presence of PA on high concentration Ca2+ aqueous subphases. Non-equilibrium relaxation (NER) experiments were also conducted and monitored by Brewster angle microscopy (BAM) to determine the effect of the Ca2+ ions on PA stability. At high surface pressures, the relaxation mechanisms of PA varied among the systems and were dependent on Ca2+ concentration.

Keyword: energy

Proteomic profiling and integrated analysis with transcriptomic data bring new insights in the stress responses of after an arrest during high-temperature ethanol fermentation.

The thermotolerant yeast is a potential candidate for high-temperature fermentation. When was used for high-temperature ethanol fermentation, a fermentation arrest was observed during the late fermentation stage and the stress responses have been investigated based on the integration of RNA-Seq and metabolite data. In order to bring new insights into the cellular responses of after the fermentation arrest during high-temperature ethanol fermentation, quantitative proteomic profiling and integrated analysis with transcriptomic data were performed in this study.Samples collected at 14, 16, 18, 20 and 22\xa0h during high-temperature fermentation were subjected to isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic profiling and integrated analysis with transcriptomic data. The correlations between transcripts and proteins for the comparative group 16\xa0h vs 14\xa0h accounted for only 4.20% quantified proteins and 3.23% differentially expressed proteins\xa0(DEPs), respectively, much higher percentages of correlations (30.56%-59.11%) were found for other comparative groups (i.e., 18\xa0h vs 14\xa0h, 20\xa0h vs 14\xa0h, and 22\xa0h vs 14\xa0h). According to Spearman correlation tests between transcriptome and proteome (the absolute value of a correlation coefficient between 0.5 and 1 indicates a strong correlation), poor correlations were found for all quantified proteins ( = -\u20090.0355 to 0.0138), DEPs ( = -\u20090.0079 to 0.0233) and the DEPs with opposite expression trends to corresponding differentially expressed genes (DEGs) ( = -\u20090.0478 to 0.0636), whereas stronger correlations were observed in terms of the DEPs with the same expression trends as the correlated DEGs ( = 0.5593 to 0.7080). The results of multiple reaction monitoring (MRM) verification indicate that the iTRAQ results were reliable. After the fermentation arrest, a number of proteins involved in transcription, translation, oxidative phosphorylation and fatty metabolism were down-regulated, some molecular chaperones and proteasome proteins were up-regulated, the ATPase activity significantly decreased, and the total fatty acids gradually accumulated. In addition, the contents of , oleic , C16, C18, C22 and C24 fatty acids increased by 16.77%, 28.49%, 14.14%, 26.88%, 628.57% and 125.29%, respectively.This study confirmed some biochemical and enzymatic alterations provoked by the stress conditions in the specific case of : such as decreases in transcription, translation and oxidative phosphorylation, alterations in cellular fatty composition, and increases in the abundance of molecular chaperones and proteasome proteins. These findings provide potential targets for further metabolic engineering towards improvement of the stress tolerance in .

Keyword: energy

Integration of proteomics and metabolomics data of early and middle season Hass avocados under heat treatment.

Ripening heterogeneity of Hass avocados results in inconsistent quality fruit delivered to the triggered and ready to eat markets. This research aimed to understand the effect of a heat shock (HS) prior to controlled atmosphere (CA) storage on the reduction of ripening heterogeneity. HS prior to CA storage reduces more drastically the ripening heterogeneity in middle season fruit. Via correlation network analysis we show the different metabolomics networks between HS and CA. High throughput proteomics revealed 135 differentially expressed proteins unique to middle season fruit triggered by HS. Further integration of metabolomics and proteomics data revealed that HS reduced the glycolytic throughput and induced protein degradation to deliver for the alternative ripening pathways. l-isoleucine, l-valine, l-aspartic and ubiquitin carboxyl-terminal hydrolase involved in protein degradation were positively correlated to HS samples. Our study provides new insights into the effectiveness of HS in synchronizing ripening of Hass avocados.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: energy

Nutritive Value of Desert Truffles Species of Genera Terfezia and Picoa (Ascomycetes) from Arid and Semiarid Regions of Eastern Turkey.

Terfezia and Picoa species contained 63-94 kcal , 76.93-83.81 g moisture, 0.78-1.52 g ash, 2.19-4.69 g protein, 2.14-9.48 g carbohydrate, 2.54-11.23 g dietary fiber, and 0.96-3.40 g fat per 100 g wet weight. We determined that Terfezia species contain more vitamin E and malondialdehyde than do Picoa species, but the amounts of vitamin A, vitamin C, and β-carotene may vary. Picoa species contain less and stearic acids but more oleic than do Terfezia species. Amino analyses revealed that glutamic and aspartic were the most abundant. We also noted that Picoa species have larger amounts of glucose and fructose than do Terfezia species. In addition, the elements these mushrooms contain can vary, but they are found at nutritious levels and are below toxic levels.

Keyword: energy

The Differentiation of Spinal Cord Motor Neurons is Associated with Changes of the Mitochondrial Phospholipid Cardiolipin.

Motor neuron damage caused by diseases, traumatic insults or de-afferentation of the spinal cord is often incurable due to the poor intrinsic regenerative capacity. Moreover, regenerated peripheral nerves often do not reach normal functionality. Here, we investigated cardiolipin in the process of neuro-differentiation, since cardiolipin is closely linked to the mitochondrial supply in cells. The NSC-34 hybrid cell line, produced by fusing neuroblastoma cells with primary spinal cord motor neurons, was used, since it shares several morphological and physiological characteristics with mature primary motor neurons. Their neuro-differentiation was supported by switching from normal to differentiation medium or by fatty supplementation. Differentiation was evaluated by measuring neurite-sprouting parameters and PPARα expression. Cellular fatty distribution was analyzed to indicate changes in lipid metabolism during differentiation. Cardiolipin was characterized by acyl-chain composition and the distribution of molecular cardiolipin species. Both, the switch from normal to differentiation medium as well as the administration of and oleic promoted neuro-differentiation. Stimulated differentiation was accompanied by changes in cardiolipin content and composition. The positive correlation between neuro-differentiation and concentration of those molecular cardiolipin species containing and oleic implied a link between differentiation of NSC-34 cells and cardiolipin metabolism. We further demonstrated the impact of cellular lipid metabolism, and particularly cardiolipin metabolism, during and NSC-34 neuritogenesis. Thus, cardiolipin may represent a new therapeutic target for axon regeneration after peripheral nerve injuries or when axon sprouting is required to compensate for motor neuron loss in response to aging and/or disease.Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.

Keyword: energy

Hemin Improves Insulin Sensitivity and Lipid Metabolism in Cultured Hepatocytes and Mice Fed a High-Fat Diet.

Hemin is a breakdown product of hemoglobin. It has been reported that the injection of hemin improves lipid metabolism and insulin sensitivity in various genetic models. However, the effect of hemin supplementation in food on lipid metabolism and insulin sensitivity is still unclear, and whether hemin directly affects cellular insulin sensitivity is yet to be elucidated. Here we show that hemin enhances insulin-induced phosphorylation of insulin receptors, Akt, Gsk3β, FoxO1 and cytoplasmic translocation of FoxO1 in cultured primary hepatocytes under insulin-resistant conditions. Furthermore, hemin diminishes the accumulation of triglyceride and increases in free fatty content in primary hepatocytes induced by palmitate. Oral administration of hemin decreases body weight, intake, blood glucose and triglyceride levels, and improves insulin and glucose tolerance as well as hepatic insulin signaling and hepatic steatosis in male mice fed a high-fat diet. In addition, hemin treatment decreases the mRNA and protein levels of some hepatic genes involved in lipogenic regulation, fatty synthesis and storage, and increases the mRNA level and enzyme activity of CPT1 involved in fatty oxidation. These data demonstrate that hemin can improve lipid metabolism and insulin sensitivity in both cultured hepatocytes and mice fed a high-fat diet, and show the potential beneficial effects of hemin from food on lipid and glucose metabolism.

Keyword: energy

Utilization of milk fatty acids by the suckling Iberian piglets.

A total of 16 pure-bred Iberian (IB) sows, all of them suckling six piglets, were used, eight of them in each of the two consecutive trials (1 and 2). Daily milk yield and composition were determined weekly over a 34-day lactation period. Within each litter, one piglet at birth and four piglets on day 35 of life were slaughtered. Milk intake per piglet tended to be greater in trial 2 (832 v. 893 g/day; P=0.066), but piglets grew at 168±3.3 g/day, irrespective of the trial. In the IB sow milk, the linoleic (LA) : linolenic (LNA) ratio averaged 14.6 and 15.2 in trial 1 and trial 2, respectively. A fivefold increase in piglet body fat content was observed over lactation (P<0.001). Most of this fat (81.4%) was present in the carcass. After 34 days of lactation, whole-body relative content of , palmitoleic, stearic and oleic acids were very close to those in the milk consumed, suggesting direct deposition. Daily deposition of LA derivatives and of LNA and its derivatives was found to be extremely low (<0.02 g, on average). Moreover, some of the arachidonic (ARA) in tissues of the IB piglet at birth disappeared throughout the lactating period. An overall fractional deposition for total fatty acids (FA) was 0.409. Fractional oxidation (disappearance) rates were 0.939 and 0.926 for n-6 and n-3 polyunsaturated FA. The overall rate of disappearance for the major non-essential FA (myristic, , palmitoleic, stearic and oleic acids), estimated as 1-the overall fractional deposition rate, was 0.546. It is concluded that the high degree of FA unsaturation, high oxidation rate of LA and LNA, and poor synthesis of ARA from LA and of docosahexaenoic from LNA found in the suckling piglet might increase the cost of whole-body fat accretion, a contributor to the observed low efficiency of use of milk for growth.

Keyword: energy

Milk production and nutrient digestibility responses to triglyceride or fatty supplements enriched in .

The objective of our study was to evaluate the effects of feeding triglyceride and fatty (FA) supplements enriched in (PA; C16:0) on production and nutrient digestibility responses of mid-lactation dairy cows. Fifteen Holstein cows (137 ± 49 d in milk) were randomly assigned to a treatment sequence in a 3 × 3 Latin square design. Treatments consisted of a control diet (CON; no added PA) or 1.5% FA added as either a FA supplement (PA-FA) or a triglyceride supplement (PA-TG). The PA supplements replaced soyhulls, and diets were balanced for glycerol content. Periods were 21 d in length with sample and data collection occurring during the final 5 d. Compared with CON, PA treatments increased dry matter (66.5 vs. 63.9%) and neutral detergent fiber (NDF) apparent digestibility (42.0 vs. 38.2%). Although PA treatments tended to increase 18-carbon FA apparent digestibility (79.1 vs. 77.9%), PA treatments decreased 16-carbon (63.1 vs. 75.8%) and total FA (72.0 vs. 76.5%) apparent digestibilities compared with CON. The PA treatments increased milk fat content (3.60 vs. 3.41%), milk fat yield (1.70 vs. 1.60 kg/d), yield of 16-carbon milk FA (570 vs. 471 g/d), 3.5% fat-corrected milk (47.6 vs. 46.5 kg/d), and -corrected milk (47.4 vs. 46.6 kg/d) compared with CON. The PA treatments did not affect dry matter intake (28.5 vs. 29.2 kg/d), milk yield (47.0 vs. 47.4 kg/d), milk protein yield (1.42 vs. 1.45 kg/d), milk lactose yield (2.29 vs. 2.31 kg/d), yield of <16-carbon milk FA (360 vs. 370 g/d), yield of >16-carbon milk FA (642 vs. 630 g/d), body weight (720 vs. 723 kg), or body condition score (3.14 vs. 3.23). We did not observe differences in digestibilities of dry matter, NDF, and 18-carbon FA between PA-TG and PA-FA. In contrast, PA-FA increased 16-carbon (68.6 vs. 57.6%) and total FA apparent digestibility (73.8 vs. 70.1%) compared with PA-TG. This resulted in PA-FA supplementation increasing the apparent digestibility of the PA supplement by ∼10 percentage points compared with PA-TG. Compared with PA-TG, PA-FA increased 16-carbon FA intake by 60 g/d, absorbed 16-carbon FA by 86 g/d, and absorbed total FA by 85 g/d. Compared with PA-TG, PA-FA increased dry matter intake (29.1 vs. 27.8 kg/d), yield of 16-carbon milk FA (596 vs. 545 g/d), and tended to increase milk yield (47.6 vs. 46.4 kg/d), milk fat yield (1.70 vs. 1.66 kg/d), and 3.5% fat-corrected milk (48.1 vs. 47.2 kg/d). In conclusion, the production response of dairy cows to PA tended to be greater for a FA supplement compared with a triglyceride supplement. Overall, PA increased NDF digestibility, milk fat yield, -corrected milk, and feed efficiency in mid-lactation dairy cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Phthalate exposure and childhood overweight and obesity: Urinary metabolomic evidence.

Metabolomics may unravel global metabolic changes in response to environmental exposures and identify important biological pathways involved in the pathophysiology of childhood obesity. Phthalate has been considered an obesogen and contributing to overweight and obesity in children. The purpose of this study is to evaluate changes in urine metabolites in response to the environmental phthalate exposure among overweight or obese children, and to investigate the metabolic mechanisms involved in the obesogenic effect of phthalate on children at puberty.Within the national Puberty Timing and Health Effects in Chinese Children (PTHEC) study, 69 overweight/obese children and 80 normal weight children were selected into the current study according to their puberty timing and WGOC (The Working Group for obesity in China) references. Urinary concentrations of six phthalate monoesters (MMP, MEP, MnBP, MEHP, MEOHP and MEHHP) were measured using API 2000 electrospray triple quadrupole mass spectrometer (ESIMS/MS). Metabolomic profiling of spot urine was performed using gas chromatography-mass spectrometry. Differentially expressed urinary metabolites associated with phthalate monoesters exposure were examined using orthogonal partial least square-discriminant analysis and multiple linear regression models. In addition, the candidate metabolites were regressed to obesity indices with multiple linear regression models and logistic regression models in all subjects.Compared with normal weight children, higher levels of MnBP were detected in urinary samples of children with overweight and obesity. After adjusting for confounders including chronological age, gender, puberty onset, daily intake and physical activity and socio-economic level, positive association remained between urinary MnBP concentration and childhood overweight/obesity [OR\u202f=\u202f1.586, 95% CI:1.043,2.412]. We observed elevated MnBP concentration was significantly correlated with increased levels of monostearin, 1-monopalmitin, stearic , itaconic , glycerol 3-phosphate, 5-methoxytryptamine, kyotorphin, 1-methylhydantoin, d-alanyl-d-alanine, pyrrole-2-carboxylic , 3,4-Dihydroxyphenylglycol, and butyraldehyde. Meanwhile, increased MnBP concentration was also significantly correlated with decreased levels of lactate, glucose 6-phosphate, d-fructose 6-phosphate, , 4-acetamidobutyric , l-glutamic , n-acetyl-l-phenylalanine, iminodiacetic , hydroxyproline, pipecolinic , l-ornithine, n-acetyl-l-glutamic , guanosine, cytosin, and (s)-mandelic in the normal weight subjects. The observations indicated that MnBP exposure was related to global urine metabolic abnormalities characterized by disrupting arginine and proline metabolism and increasing oxidative stress and fatty reesterification. Among the metabolic markers related to MnBP exposure, 1-methylhydantoin, pyrrole-2-carboxylic and monostearin were found to be positively correlated with obesity indices, while hydroxyproline, l-ornithine, and lactate were negatively associated with overweight/obesity in children.Our results suggested that the disrupted arginine and proline metabolism associated with phthalate exposure might contribute to the development of overweight and obesity in school-age children, providing insights into the pathophysiological changes and molecular mechanisms involved in childhood obesity.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Efficacy of bezafibrate on fibroblasts of glutaric acidemia type II patients evaluated using an in vitro probe acylcarnitine assay.

We evaluated the effects of bezafibrate (BEZ) on β-oxidation in fibroblasts obtained from patients with glutaric acidemia type II (GA2) of various clinical severities using an in vitro probe (IVP) assay.Cultured fibroblasts from 12 patients with GA2, including cases of the neonatal-onset type both with and without congenital anomalies (the prenatal- and neonatal-onset forms, respectively), the infantile-onset, and the myopathic forms, were studied. The IVP assay was performed by measuring acylcarnitines (ACs) in the cell culture medium of fibroblasts incubated with for 96h in the presence of 0-800μM BEZ using tandem mass spectrometry.The IVP assay showed that 100μM BEZ markedly reduced the level of palmitoylcarnitine (C16) in the neonatal-onset, infantile-onset, and myopathic forms of GA2, either increasing or maintaining a high level of acetylcarnitine (C2), which serves as an index of production via β-oxidation. In the prenatal-onset form, although a small reduction of C16 was also observed in the presence of 100μM BEZ, the level of C2 remained low. At concentrations higher than 100μM, BEZ further decreased the level of ACs including C16, but a concentration over 400μM decreased the level of C2 in most cases.BEZ at 100μM was effective for all GA2 phenotypes except for the prenatal-onset form, as a reduction of C16 without deterioration of C2 is considered to indicate improvement of β-oxidation. The effects of higher doses BEZ could not be estimated by the IVP assay but might be small or nonexistent.Copyright © 2016 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.

Keyword: energy

Antibacterial activity of extracted bioactive molecules of Schinus terebinthifolius ripened fruits against some pathogenic bacteria.

The aim of this work is to identify the chemical constituents and the bioactivity of essential oil (EO), acetone extract (ACE) and n-hexane extract (HexE) of S. terebinthifolius ripened fruits using GC-MS. Total phenolic content and antioxidant activity of extracts were determined using the Folin-Ciocalteu and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assays, respectively. The toxicity against the growth of Acinetobacter baumannii, Bacillus subtilis, Escherichia coli, Micrococcus flavus, Pseudomonas aeruginosa, Sarcina lutea, and Staphylococcus aureus was determined with measuring the inhibition zones (IZs) using the disc diffusion method at the concentrations from 125 to 2000\u202fμg/mL, also, the minimum inhibitory concentrations (MICs) using 96-well micro-plates and ranged from 4 to 2000\u202fμg/mL. The major components in EO were α-pinene (36.9%), and α-phellandrene (32.8%). The major components in ACE were oleic (38.7%), α-phellandrene (13.33%), and δ-cadinene (11.1%), while the major methyl esters of fatty acids detected in HexE were oleic (12.8%), and (10.9%). The EO showed good activity against the growth of Staph. aureus and P. aeruginosa with MIC values of 16\u202fμg/mL and 32\u202fμg/mL, the ACE showed broad activity against the studied bacterial pathogens with MIC values ranged from of 4-128\u202fμg/mL against the studied bacterial isolates, while HexE, however, showed weak antibacterial activity. The IC values of EO, ACE and HexE were 15.11\u202f±\u202f0.99, 118.16\u202f±\u202f1.7 and 324.26\u202f±\u202f2.45\u202fμg/mL, respectively, compared to IC of Tannic (23.83\u202f±\u202f1.9\u202fμg/mL) and butylated hydroxytoluene (BHT, 2.9\u202f±\u202f0.1\u202fμg/mL). Data suggested that the ripened fruits of S. terebinthifolius have potent antioxidant and antibacterial activities.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Superhydrophobic Polypropylene Functionalized with Nanoparticles for Efficient Fast Static and Dynamic Separation of Spilled Oil from Water.

Frequent oil spills not only threaten the ecosystem, but they are also a waste of a valuable source of . There is an urgent need to develop materials that can readily remove spilled oil from water bodies and also have the capacity to collect it for applications. Herein, a superhydrophobic fiber of functionalized polypropylene is engineered with the help of interaction with incorporated copper oxide nanoparticles. The successful development of functionalized polypropylene is confirmed by Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, and -dispersive X-ray spectroscopy. The scanning electron microscopy images reveal that the surface roughness of the polypropylene is enhanced after functionalization. The optimized functionalized polypropylene displays an ultrahydrophobic surface with a water contact angle of 162.42°. The functionalized polyprolyene displays good absorption capacity. It has the capacity to take 30 to 40 times its own weight in oils and nonpolar organic solvents, which makes it useful for small spills. With a flux of 11 204 Lm h, functionalized polypropylene is as an ideal material for the dynamic separation of oil spills from water. It also has excellent selectivity towards oil, water rejection, and oil absorption capacity.© 2019 The Authors. Published by WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: energy

[THE UNESTERIFIED FATTY ACIDS IN BLOOD PLASMA AND INTERCELLULAR MEDIUM: EFFECT OF INSULIN AND ALBUMIN (THE LECTURE)].

The high content of saturated fatty , triglycerides in food, the large amount of lipoproteins of very low density of the same name in blood, obvious insufficient amount of unesterified fatty acids releasing under lipolysis in blood to meet in vivo requirements in biotransforming of ATP are the causes of biological malfunction of homeostasis. As a rule, for every cell in vivo everything is always to be enough. The deficiency of synthesis of ATP by reason of non-optimal substratum for acquirement of ATP by mitochondria is followed by activation also phylogenetically earlier biological function of adaptation, biological reaction of stress. Thus, surplus of unesterified fatty after every food intake forms in vivo biological reaction of "metabolic" stress, deficiency of by reason of realization by mitochondria in vivo non-optimal exogenous substratum- unesterified fatty , deficiency of acyl- and acetyl-KoA and prognostically formation of potentially ineffective alternative of metabolism of fatty acids. The deficiency of unesterified fatty acids in biological reaction of exotrophy after every intake of food compensates biological reaction of stress, activation of releasing of unesterified fatty acids from visceral fatty cells of gland as it physiologically occurs in biological reaction of endotrophy. At that, adrenalin increases lipolysis in visceral fatty cells of gland and physiologically late insulin can\'t inhibit lipolysis in phylogenetically early visceral fatty cells. Increasing of content of unesterifed fatty acids in blood plasma, as it always occurs in vivo, stops absorption of glucose by cells initiating hyperglycemia, hyperinsulinemia, and syndrome of resistance to insulin. The result of such a compensation of biological reaction of exotrophy is biological reaction of endotrophy, condition of "metabolic" stress, depletion of function of β-cells of islets with formation of diabetes mellitus type I, deficiency in vivo of insulin synthesis. The biological role of albumin - transfer of fatty acids in intercellular medium inform of unesterifed fatty acids and prevention of formation of pool of free fatty acids effecting aphysiologically.

Keyword: energy

Microalgae Characterization for Consolidated and New Application in Human Food, Animal Feed and Nutraceuticals.

The exploration of new food sources and natural products is the result of the increase in world population as well as the need for a healthier diet; in this context, microalgae are undoubtedly an interesting solution. With the intent to enhance their value in new commercial applications, this paper aims to characterize microalgae that have already been recognized as safe or authorized as additives for humans and animals (, , , ) as well as those that have not yet been marketed ( and sp.). In this scope, the content of proteins, carbohydrates, lipids, total dietary fiber, humidity, ash, and carotenoids has been measured via standard methods. In addition, individual carotenoids (beta-carotene, astaxanthin, and lutein) as well as individual saturated, monounsaturated, and polyunsaturated fatty acids have been identified and quantified chromatographically. The results confirm the prerogative of some species to produce certain products such as carotenoids, polyunsaturated fatty acids, and proteins, but also show how their cellular content is rich and diverse. green and red phases, and sp., in addition to producing astaxanthin and omega-3, contain about 25⁻33% / proteins on a dry basis. is rich in beta-carotene (3.45% / on a dry basis), is a source of lutein (0.30% / on a dry basis), and the species is a protein-based microalgae (45% / on a dry basis). All, however, can also produce important fatty acids such as , γ-linolenic , and oleic . Considering their varied composition, these microalgae can find applications in multiple sectors. This is true for microalgae already on the market as well as for promising new sources of bioproducts such as and sp.

Keyword: energy

Central composite design parameterization of microalgae/cyanobacteria co-culture pretreatment for enhanced lipid extraction using an external clamp-on ultrasonic transducer.

Lipids extracted from algal biomass could provide an abundant, rapidly growing, high yield feedstock for bio-diesel and other green fuels to supplement current fossil-based sources. Ultrasound pretreatment is a mechanical cell disruption method that has been shown to enhance lipid recovery from algae due to cavitation effects that disrupt algae cell walls. In this study, a locally grown mixture of Chlorella vulgaris/Cyanobacteria leptolyngbya was sonicated in an ultrasonic reactor with a clamp-on transducer prior to solvent lipid extraction. This configuration allows for a non-contact delivery method of ultrasonic with improved operational advantages (no fouling of transducer, continuous operation, and fully scalable design). A central composite design (CCD) was implemented to statistically analyze and evaluate the effect of ultrasonic power (350-750\u202fW) and treatment time (5-30\u202fmin) on lipid yield. Lipid recovery was found to increase with both ultrasonic power and treatment time. Total lipid yields (on dry biomass basis) extracted via the Bligh and Dyer method from Chlorella vulgaris/cyanobacteria co-culture ranged from 8.3% for untreated algae to 16.9% for algae sonicated with 750\u202fW power for 30\u202fmin, which corresponds to more than a doubling of lipid recovery due to ultrasound pretreatment. Increased power and treatment times were found to increase the degree of cell disruption as observed in the SEM and TEM images after ultrasonic pretreatment. Additionally, hexane (1:1 v/v) was evaluated as an alternative to the standard Bligh & Dyer (2:2:1.8 v/v/v chloroform/methanol/cell suspension) lipid extraction solvent system. On average, the Bligh and Dyer method extracted on average over twice the amount of lipids compared to hexane extraction. The lipid profile of the algae extracts indicates high concentrations of lauric (12:0), (16:0), stearic (18:0), oleic (18:1), and linoleic (18:2). This particular configuration of an ultrasonic system proved to be a viable method for the pretreatment of algae for enhanced lipid yields. Future research should focus on identifying alternative extraction solvents and expanding the range of treatment conditions to optimize the ultrasonic power and treatment times for maximum lipid recovery.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

is a novel hypothalamic gene upregulated by a high-fat diet and leptin in mice.

homeostasis is regulated by the hypothalamus but fails when animals are fed a high-fat diet (HFD), and leptin insensitivity and obesity develops. To elucidate the possible mechanisms underlying these effects, a microarray-based transcriptomics approach was used to identify novel genes regulated by HFD and leptin in the mouse hypothalamus.Mouse global array data identified as a novel gene highly upregulated by both a HFD and leptin challenge. In situ hybridisation showed expression upregulation by HFD and leptin in all major hypothalamic nuclei in agreement with transcriptomic gene expression data. Immunohistochemistry and studies in the hypothalamic clonal neuronal cell line, mHypoE-N42 (N42), confirmed that alpha 1-antichymotrypsin (αAC), the protein encoded by , is localised to neurons and revealed that it is secreted into the media. expression in N42 neurons is upregulated by and by leptin, together with and , and all three genes are downregulated by the anti-inflammatory monounsaturated fat, oleic . Additionally, palmitate upregulation of in N42 neurons is blocked by the NFκB inhibitor, BAY11, and the upregulation of expression in the hypothalamus by HFD is blunted in IL-1 receptor 1 knockout ( ) mice.These data demonstrate that expression is implicated in nutritionally mediated hypothalamic inflammation.

Keyword: energy

Lipids in human milk.

Fat is the main providing component in human milk and comprising a complex mixture of different lipid species, with quantitative dominance of triglycerides. After elucidating the fatty composition, more recent research has looked at influencing factors and the importance of specific lipids. Here we review quantitative aspects of maternal metabolism which contribute to the milk fatty composition, especially considering essential fatty acids and their long chain polyunsaturated derivatives. In this context studies with stable isotopes have indicated the importance of maternal body pools for mediating the effects of diet on milk composition. Furthermore, the importance of positioning of at the\xa0glycerol backbone of triglycerides is discussed, and the phospholipids of the milk fat globule membrane are described and examples for their potential importance for infant development are presented.Copyright © 2017 Elsevier Ltd. All rights reserved.

Keyword: energy

New compounds inhibiting lipid accumulation from the stems of Sabia parviflora.

Four new compounds, sabianin A (1), sabianin B (2), sabianin C, (3) and sabianin D (4) together with seven known compounds (5-11) were isolated from the stems of Sabia parviflora. The structures of these compounds were elucidated by extensive spectroscopic analysis and by comparing their NMR data with those of related compounds. Their absolute configurations were determined by comparing the calculated and experimental electronic circular dichroism spectra. The lipid-lowering effect of all the compounds was evaluated using a cell model of steatosis induced by oleic and . Compounds 3, 6, and 10 showed statistically significant lipid-lowering effect.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: energy

Effect of Hyperbaric Oxygen Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Diabetes Mellitus Patients: A Pilot Study.

Metabolic changes in insulin-dependent diabetes mellitus (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the lipid profile and altering FA composition.Copyright © 2019 Canadian Diabetes Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Effect of flue gas CO on the growth, carbohydrate and fatty composition of a green microalga Scenedesmus obliquus for biofuel production.

Effect of various flue gas CO concentrations (5%, 10% and 14.1%) on growth rate and biochemical properties of a green microalga Scenedesmus obliquus was investigated. S. obliquus showed the highest biomass production and growth rate (0.36\u2005g\u2005L and μ\u2009=\u20091.00\u2005day), total inorganic carbon removal (35.8\u2005mg\u2005L), lipid productivity (9.9\u2005mg\u2005L\u2005day) and carbohydrate productivity (10.3\u2005mg\u2005L\u2005day) with 14.1% CO after 8 days of cultivation. Fatty methyl ester analysis revealed that the and oleic contents were increased up to 5% and 7% with 14.1% CO, respectively. Application of flue gas CO enhanced the growth along with lipid and carbohydrate productivity of S. obliquus, which can be exploited for reducing the CO concentration.

Keyword: energy

Effect of on the mitigation of milk fat depression syndrome caused by trans-10, cis-12-conjugated linoleic in grazing dairy cows.

The objective of the study was to evaluate the effect of adding protected (PA) to the ration of grazing dairy cows supplemented with protected conjugated linoleic (CLA) on milk production, chemical composition and fat profile. Six cows were used, 3/4 American Swiss\xa0×\xa0Zebu, under a rotational grazing system in a mixed sward with Cynodon plectostachyus, Brachiaria decumbens and Brachiaria brizantha. Furthermore, each cow received daily 4\xa0kg concentrates and 8\xa0kg sorghum silage, which made up the basal diet. The cows were distributed into three two-cow groups. Three treatments were randomly assigned to the groups, using a cross design: (1) control (basal diet), (2) basal diet\xa0+\xa0CLA (50\xa0g/d) and (3) basal diet\xa0+\xa0CLA (50\xa0g/d)\xa0+\xa0PA (412\xa0g/d). The following variables were evaluated: forage intake, milk production, protein, fat and lactose concentration in milk, and milk fatty (FA) profile. There were no differences in forage intake between treatments; however, there were differences in milk production, protein, fat and lactose yield and fat concentration, which increased significantly in group CLA\xa0+\xa0PA when compared with group CLA. The concentration of FA synthesised de novo was lower when PA was included in the diet. Adding PA to the diet of grazing cows mitigates the milk fat decline caused by including trans-10, cis-12 CLA in the diet.

Keyword: energy

Impact of Maternal Diet on Human Milk Composition Among Lactating Women in Latvia.

: Many studies indicate that the maternal diet is an important factor affecting human milk composition. Human milk composition among lactating women in Latvia, as well as the maternal diet during lactation, has not been sufficiently studied. The aim of this research was to assess dietary habits and macronutrient intake among lactating women in Latvia and to examine the effect of diet on human milk composition. : Research was conducted between November 2016 and December 2017. Mature human milk samples ( = 61) along with a 72h food diary, a food frequency questionnaire (FFQ), and a questionnaire about maternal and infant characteristics were obtained from voluntary women who were recruited via an invitation published in a social media member group for nursing mothers. Fat content in human milk was determined by LVS ISO 2446:2008, protein content was determined by LVS EN ISO 8968-1:2014, lactose was determined by ISO 22662:2007, and the fatty profile was analyzed using gas chromatography. Dietary data were evaluated using the Finnish food composition database Fineli, release 19 (3 March 2018). : Median values for fat, protein, and lactose in mature human milk were 4.40%, 1.08%, and 6.52%, respectively. Predominant fatty acids in human milk were oleic (C18:1 n9), (C16:0), and linoleic (C18:2 n6) at 34.60%, 24.00%, and 11.00% of total fatty acids, respectively. The elaidic (C18:1 n9) level was <0.10% in all human milk samples. Significant, positive associations ( < 0.05) were found between maternal dietary intake of linoleic, α-linolenic, docosahexaenoic, total -monounsaturated, total -polyunsaturated, and total n-6 and n-3 polyunsaturated fatty acids, the ratio of n-6/n-3, and the level of these fatty acids in human milk. Total and carbohydrate intake among participants were lower, but total fat, saturated fat, and sugar intake were higher than recommended. Protein, linoleic , and α-linolenic intake were adequate, but docosahexaenoic intake was noticeably lower than recommended. Women should be supported with information regarding their nutritional needs during lactation and the possible impact of diet on human milk composition. : Macronutrient (fat, protein, and lactose) content in human milk is not affected by maternal diet. Conversely, the human milk fatty profile is affected by the immediate diet consumed by the mother. Habitual dietary habits can also impact the fatty profile of human milk.

Keyword: energy

Intake of individual saturated fatty acids and risk of coronary heart disease in US men and women: two prospective longitudinal cohort studies.

\xa0To investigate the association between long term intake of individual saturated fatty acids (SFAs) and the risk of coronary heart disease, in two large cohort studies.\xa0Prospective, longitudinal cohort study.\xa0Health professionals in the United States.\xa073\u2009147 women in the Nurses\' Health Study (1984-2012) and 42\u2009635 men in the Health Professionals Follow-up Study (1986-2010), who were free of major chronic diseases at baseline.\xa0Incidence of coronary heart disease (n=7035) was self-reported, and related deaths were identified by searching National Death Index or through report of next of kin or postal authority. Cases were confirmed by medical records review.\xa0Mean intake of SFAs accounted for 9.0-11.3% intake over time, and was mainly composed of lauric (12:0), myristic (14:0), (16:0), and stearic (18:0; 8.8-10.7% ). Intake of 12:0, 14:0, 16:0 and 18:0 were highly correlated, with Spearman correlation coefficients between 0.38 and 0.93 (all P<0.001). Comparing the highest to the lowest groups of individual SFA intakes, hazard ratios of coronary heart disease were 1.07 (95% confidence interval 0.99 to 1.15; P=0.05) for 12:0, 1.13 (1.05 to 1.22; P<0.001) for 14:0, 1.18 (1.09 to 1.27; P<0.001) for 16:0, 1.18 (1.09 to 1.28; P<0.001) for 18:0, and 1.18 (1.09 to 1.28; P<0.001) for all four SFAs combined (12:0-18:0), after multivariate adjustment of lifestyle factors and total intake. Hazard ratios of coronary heart disease for isocaloric replacement of 1% from 12:0-18:0 were 0.92 (95% confidence interval 0.89 to 0.96; P<0.001) for polyunsaturated fat, 0.95 (0.90 to 1.01; P=0.08) for monounsaturated fat, 0.94 (0.91 to 0.97; P<0.001) for whole grain carbohydrates, and 0.93 (0.89 to 0.97; P=0.001) for plant proteins. For individual SFAs, the lowest risk of coronary heart disease was observed when the most abundant SFA, 16:0, was replaced. Hazard ratios of coronary heart disease for replacing 1% from 16:0 were 0.88 (95% confidence interval 0.81 to 0.96; P=0.002) for polyunsaturated fat, 0.92 (0.83 to 1.02; P=0.10) for monounsaturated fat, 0.90 (0.83 to 0.97; P=0.01) for whole grain carbohydrates, and 0.89 (0.82 to 0.97; P=0.01) for plant proteins.\xa0Higher dietary intakes of major SFAs are associated with an increased risk of coronary heart disease. Owing to similar associations and high correlations among individual SFAs, dietary recommendations for the prevention of coronary heart disease should continue to focus on replacing total saturated fat with more healthy sources of .Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Keyword: energy

Attenuation of Free Fatty -Induced Muscle Insulin Resistance by Rosemary Extract.

Elevated blood free fatty acids (FFAs), as seen in obesity, impair muscle insulin action leading to insulin resistance and Type 2 diabetes mellitus. Serine phosphorylation of the insulin receptor substrate (IRS) is linked to insulin resistance and a number of serine/threonine kinases including JNK, mTOR and p70 S6K have been implicated in this process. Activation of the sensor AMP-activated protein kinase (AMPK) increases muscle glucose uptake, and in recent years AMPK has been viewed as an important target to counteract insulin resistance. We reported recently that rosemary extract (RE) increased muscle cell glucose uptake and activated AMPK. However, the effect of RE on FFA-induced muscle insulin resistance has never been examined. In the current study, we investigated the effect of RE in palmitate-induced insulin resistant L6 myotubes. Exposure of myotubes to palmitate reduced the insulin-stimulated glucose uptake, increased serine phosphorylation of IRS-1, and decreased the insulin-stimulated phosphorylation of Akt. Importantly, exposure to RE abolished these effects and the insulin-stimulated glucose uptake was restored. Treatment with palmitate increased the phosphorylation/activation of JNK, mTOR and p70 S6K whereas RE completely abolished these effects. RE increased the phosphorylation of AMPK even in the presence of palmitate. Our data indicate that rosemary extract has the potential to counteract the palmitate-induced muscle cell insulin resistance and further studies are required to explore its antidiabetic properties.

Keyword: energy

Disentangling the photochemical salinity tolerance in Aster tripolium L.: connecting biophysical traits with changes in fatty composition.

A profound analysis of A.\xa0tripolium photochemical traits under salinity exposure is lacking in the literature, with very few references focusing on its fatty profile role in photophysiology. To address this, the deep photochemical processes were evaluated by Pulse Amplitude Modulated (PAM) Fluorometry coupled with a discrimination of its leaf fatty profile. Plants exposed to 125-250\xa0mm NaCl showed higher photochemical light harvesting efficiencies and lower dissipation rates. under higher NaCl exposure, there is evident damage of the oxygen evolving complexes (OECs). On the other hand, Reaction Centre (RC) closure net rate and density increased, improving the fluxes entering the PS II, in spite of the high amounts of dissipated and the loss of PS II antennae connectivity. dissipation was mainly achieved through the auroxanthin pathway. Total fatty content displayed a similar trend, being also higher under 125-250\xa0mm NaCl with high levels of omega-3 and omega-6 fatty acids. The increase in oleic and allows the maintenance of the good functioning of the PS II. Also relevant was the high concentration of chloroplastic C16:1t in the individuals subjected to 125-250\xa0mm NaCl, related with a higher electron transport activity and with the organization of the Light Harvesting Complexes (LHC) and thus reducing the activation of dissipation mechanisms. All these new insights shed some light not only on the photophysiology of this potential cash-crop, but also highlight its important saline agriculture applications of this species as forage and potential source of essential fatty acids.© 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

Keyword: energy

Environmental temperature alters the overall digestive energetics and differentially affects dietary protein and lipid use in a lizard.

Processing food (e.g. ingestion, digestion, assimilation) requires referred to as specific dynamic action (SDA) and is at least partially fuelled by oxidation of the nutrients (e.g. proteins and lipids) within the recently ingested meal. In ectotherms, environmental temperature can affect the magnitude and/or duration of the SDA, but is likely to also alter the mixture of nutrients that are oxidized to cover these costs. Here, we examined metabolic rate, gut passage time, assimilation efficiency and fuel use in the lizard digesting cricket meals at three ecologically relevant temperatures (20, 25 and 32°C). Crickets were isotopically enriched with C-leucine or C- tracers to distinguish between protein and lipid oxidation, respectively. Our results show that higher temperatures increased the magnitude of the SDA peak (by 318% between 32 and 20°C) and gut passage rate (63%), and decreased the duration of the SDA response (by 20% for males and 48% for females). Peak rate of dietary protein oxidation occurred sooner than peak lipid oxidation at all temperatures (70, 60 and 31\u2005h earlier for 20, 25 and 32°C, respectively). Assimilation efficiency of proteins, but not lipids, was positively related to temperature. Interestingly, the SDA response exhibited a notable circadian rhythm. These results show that temperature has a pronounced effect on digestive energetics in , and that this effect differs between nutrient classes. Variation in environmental temperatures may thus alter the budget and nutrient reserves of these animals.© 2019. Published by The Company of Biologists Ltd.

Keyword: energy

Feeding of palm oil fatty acids or rapeseed oil throughout lactation: Effects on status, body composition, and milk production in Norwegian dairy goats.

The objective of this experiment was to examine how supplements of rapeseed oil or palm oil fatty acids would affect milk production and composition, body lipid stores, and balance in 30 multiparous goats of Norwegian dairy goat breed. The experiment lasted 230 d, with 1 to 120 d in milk (DIM) for indoor feeding (P1), 120 to 200 DIM for mountain grazing (P2), and 200 to 230 DIM for indoor feeding (P3). Grass silage was fed according to appetite during indoor feeding periods. After an adjustment period (1-60 DIM) when the control diet was given to the goats, the animals were subdivided into 3 groups of 10 goats. Treatments (60-230 DIM) were (1) basal concentrate (control; no added fat); (2) control concentrate with 8% (added on air-dry basis) hydrogenated palm oil enriched with (POFA); and (3) control concentrate with 8% (added on air-dry basis) rapeseed oil (RSO). Individual balances based on intake and milk production were estimated on 10, 30, 60, 90, 120, 200, and 230 DIM. At the same times, body weight (BW), body condition score (BCS), body mass index, and body tissue stores using computed tomography were monitored. Silage intake was depressed by POFA throughout the experimental period. Reduced BW and body mass index were observed in the POFA and RSO groups, whereas no effect on BCS or body composition was observed throughout lactation. Generally, a minor decrease in BW was observed from 10 to 120 DIM (only 0.6 kg on average) and the total amount of body lipid was reduced by 4.4 kg. During the mountain grazing period, a further reduction in body lipid stores (2.7 kg) was observed, and BW was reduced by 3.9 kg in the same period. The goats mobilized, on average, 72% of their fat reserves during the first 200 DIM. In this period, dietary fat supplementation did not reduce the mobilization of adipose tissue but resulted in greater milk fat yield (2 kg more, on average, compared with the control group). Milk yield was not affected by POFA or RSO supplementation. Milk fat content was higher in the POFA group than in the control and RSO groups. Milk protein and lactose contents were not affected by lipid supplements. In late lactation, a rapid accumulation of fat deposits followed the intense mobilization during the grazing period. Dietary lipid supplements had no effect on milk fat yield at this stage. Milk production depends heavily on the ability to mobilize body lipid stores, and neither POFA nor RSO supplements at rates used in our study affected this mobilization.Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Dihydrosterculic from cottonseed oil suppresses desaturase activity and improves liver metabolomic profiles of high-fat-fed mice.

Polyunsaturated fatty (PUFA)-rich diets are thought to provide beneficial effects toward metabolic health in part through their bioactive properties. We hypothesized that increasing PUFA intake in mice would increase peroxisome proliferator activated receptor delta (PPARδ) expression and activity, and we sought to examine the effect of different PUFA-enriched oils on muscle PPARδ expression. One of the oils we tested was cottonseed oil (CSO) which is primarily linoleic (53%) and (24%). Mice fed a CSO-enriched diet (50% from fat) displayed no change in muscle PPARδ expression; however, in the liver, it was consistently elevated along with its transcriptional coactivator Pgc-1. Male mice were fed chow or CSO-, saturated fat (SFA)-, or linoleic (18:2)-enriched diets that were matched for macronutrient content for 4 weeks. There were no differences in food intake, body weight, fasting glucose, glucose tolerance, or expenditure between chow- and CSO-fed mice, whereas SFA-fed mice had increased fat mass and 18:2-fed mice were less glucose tolerant. Metabolomic analyses revealed that the livers of CSO-fed mice closely matched those of chow-fed but significantly differed from SFA- and 18:2-enriched groups. Fatty composition of the diets and livers revealed an impairment in desaturase activity and the presence of dihydrosterculic (DHSA) in the CSO-fed mice. The effect of DHSA on PPARδ and stearoyl-CoA desaturase-1 expression mimicked that of the CSO-fed mice. Taken together, these data suggest that DHSA from CSO may be an effective means to increase PPARδ expression with concomitant suppression of liver stearoyl-CoA desaturase-1 activity.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

Multi-omic profiles of hepatic metabolism in TPN-fed preterm pigs administered new generation lipid emulsions.

We aimed to characterize the lipidomic, metabolomic, and transcriptomic profiles in preterm piglets administered enteral (ENT) formula or three parenteral lipid emulsions [parenteral nutrition (PN)], Intralipid (IL), Omegaven (OV), or SMOFlipid (SL), for 14 days. Piglets in all parenteral lipid groups showed differential organ growth versus ENT piglets; whole body growth rate was lowest in IL piglets, yet there were no differences in either expenditure or (13)C-palmitate oxidation. Plasma homeostatic model assessment of insulin resistance demonstrated insulin resistance in IL, but not OV or SL, compared with ENT. The fatty and acyl-CoA content of the liver, muscle, brain, and plasma fatty acids reflected the composition of the dietary lipids administered. Free carnitine and acylcarnitine (ACT) levels were markedly reduced in the PN groups compared with ENT piglets. Genes associated with oxidative stress and inflammation were increased, whereas those associated with alternative pathways of fatty oxidation were decreased in all PN groups. Our results show that new generation lipid emulsions directly enrich tissue fatty acids, especially in the brain, and lead to improved growth and insulin sensitivity compared with a soybean lipid emulsion. In all total PN groups, carnitine levels are limiting to the formation of ACTs and gene expression reflects the stress of excess lipid on liver function.

Keyword: energy

Impact of novel palmitoylated prolactin-releasing peptide analogs on metabolic changes in mice with diet-induced obesity.

Analogs of anorexigenic neuropeptides, such as prolactin-releasing peptide (PrRP), have a potential as new anti-obesity drugs. In our previous study, attached to the N-terminus of PrRP enabled its central anorexigenic effects after peripheral administration. In this study, two linkers, γ-glutamic at Lys11 and a short, modified polyethylene glycol at the N-terminal Ser and/or Lys11, were applied for the palmitoylation of PrRP31 to improve its bioavailability. These analogs had a high affinity and activation ability to the PrRP receptor GPR10 and the neuropeptide FF2 receptor, as well as short-term anorexigenic effect similar to PrRP palmitoylated at the N-terminus. Two-week treatment with analogs that were palmitoylated through linkers to Lys11 (analogs 1 and 2), but not with analog modified both at the N-terminus and Lys11 (analog 3) decreased body and liver weights, insulin, leptin, triglyceride, cholesterol and free fatty plasma levels in a mouse model of diet-induced obesity. Moreover, the expression of uncoupling protein-1 was increased in brown fat suggesting an increase in expenditure. In addition, treatment with analogs 1 and 2 but not analog 3 significantly decreased urinary concentrations of 1-methylnicotinamide and its oxidation products N-methyl-2-pyridone-5-carboxamide and N-methyl-4-pyridone-3-carboxamide, as shown by NMR-based metabolomics. This observation confirmed the previously reported increase in nicotinamide derivatives in obesity and type 2 diabetes mellitus and the effectiveness of analogs 1 and 2 in the treatment of these disorders.

Keyword: energy

Dynamics of Individual Fatty Acids in Muscle Fat Stores and Membranes of a Songbird and Its Functional and Ecological Importance.

Although tissue fatty (FA) composition has been linked to whole-animal performance (e.g., aerobic endurance, metabolic rate, postexercise recovery) in a wide range of animal taxa, we do not adequately understand the pace of changes in FA composition and its implications for the ecology of animals. Therefore, we used a C to C diet shift experiment and compound-specific δC analysis to estimate the turnover rates of FAs in the polar and neutral fractions of flight muscle lipids (corresponding to membranes and lipid droplets) of exercised and sedentary zebra finches (Taeniopygia guttata). Turnover was fastest for linoleic (LA; 18:2n6) and (PA; 16:0), with 95% replacement times of 10.8-17.7 d in the polar fraction and 17.2-32.8 d in the neutral fraction, but was unexpectedly slow for the long-chain polyunsaturated FAs (LC-PUFAs) arachidonic (20:4n6) and docosahexaenoic (22:6n3) in the polar fraction, with 95% replacement in 64.9-136.5 d. Polar fraction LA and PA turnover was significantly faster in exercised birds (95% replacement in 8.5-13.3 d). Our results suggest that FA turnover in intramuscular lipid droplets is related to FA tissue concentrations and that turnover does not change in response to exercise. In contrast, we found that muscle membrane FA turnover is likely driven by a combination of selective LC-PUFA retention and consumption of shorter-chain FAs in metabolism. The unexpectedly fast turnover of membrane-associated FAs in muscle suggests that songbirds during migration could substantially remodel their membranes within a single migration stopover, and this may have substantial implications for how the FA composition of diet affects metabolism of birds during migration.

Keyword: energy

High density lipoprotein (HDL) reverses induced metabolism imbalance by switching CD36 and GLUT4 signaling pathways in cardiomyocyte.

In our previous study (PA) induced lipotoxicity and switches metabolism from CD36 to GLUT4 in H9c2 cells. Low level of high density lipoprotein (HDL) is an independent risk factor for cardiac hypertrophy. Therefore, we in the present study investigated whether HDL can reverse PA induced lipotoxicity in H9c2 cardiomyoblast cells. In this study, we treated H9c2 cells with PA to create a hyperlipidemia model in vitro and analyzed for CD36 and GLUT4 metabolic pathway proteins. CD36 metabolic pathway proteins (phospho-AMPK, SIRT1, PGC1α, PPARα, CPT1β, and CD36) were decreased by high PA (150 and 200\u2009μg/μl) concentration. Interestingly, expression of GLUT4 metabolic pathway proteins (p-PI3K and pAKT) were increased at low concentration (50\u2009μg/μl) and decreased at high PA concentration. Whereas, phospho-PKCζ, GLUT4 and PDH proteins expression was increased in a dose dependent manner. PA treated H9c2 cells were treated with HDL and analyzed for cell viability. Results showed that HDL treatment induced cell proliferation efficiency in PA treated cells. In addition, HDL reversed the metabolic effects of PA: CD36 translocation was increased and reduced GLUT4 translocation, but HDL treatment significantly increased CD36 metabolic pathway proteins and reduced GLUT4 pathway proteins. Rat neonatal cardiomyocytes showed similar results. In conclusion, HDL reversed palmatic -induced lipotoxicity and metabolism imbalance in H9c2 cardiomyoblast cells and in neonatal rat cardiomyocyte cells.© 2017 Wiley Periodicals, Inc.

Keyword: energy

A new approach of microalgal biomass pretreatment using deep eutectic solvents for enhanced lipid recovery for biodiesel production.

The biomass of Chlorella sp. was pretreated with three different aqueous deep eutectic solvents (aDESs), i.e. aqueous choline chloride-oxalic (aCh-O), aqueous choline chloride-ethylene glycol (aCh-EG) and aqueous urea-acetamide (aU-A). The effect of aDESs pretreatment of microalgae biomass was evaluated in terms of lipid recovery rate, total carbohydrate content, fatty composition, and thermal chemical behavior of biomass. Results indicated that, lipid recovery rate was increased from 52.03% of untreated biomass to 80.90%, 66.92%, and 75.26% of the biomass treated by aCh-O, aCh-EG and aU-A, respectively. However, there were no major changes observed in fatty profiles of both untreated and treated biomass, specifically , palmitoleic and stearic under various pretreatments. Furthermore, characterizations of untreated and treated biomass were carried out using Fourier transform infrared (FTIR), thermogravimetry analysis (TGA) and scanning electron microscope (SEM) to understand the enhanced lipids recovery.Copyright © 2016. Published by Elsevier Ltd.

Keyword: energy

1\'-Acetoxychavicol acetate ameliorates age-related spatial memory deterioration by increasing serum ketone body production as a complementary source for neuronal cells.

1\'-Acetoxychavicol acetate (ACA) is naturally obtained from the rhizomes and seeds of Alpinia galangal. Here, we examined the effect of ACA on learning and memory in senescence-accelerated mice prone 8 (SAMP8). In mice that were fed a control diet containing 0.02% ACA for 25 weeks, the learning ability in the Morris water maze test was significantly enhanced in comparison with mice that were fed the control diet\xa0alone. In the Y-maze test, SAMP8 mice showed decreased spontaneous alterations in comparison with senescence-accelerated resistant/1 (SAMR1) mice, a homologous control, which was improved by ACA pretreatment. Serum metabolite profiles were obtained by GC-MS analysis, and each metabolic profile was plotted on a 3D score plot. Based upon the diagram, it can be seen that the distribution areas for the three groups were completely separate. Furthermore, the contents of β-hydroxybutyric and in the serum of SAMP8-ACA mice were higher than those of SAMP8-control mice and SAMR1-control mice. We also found that SAMR1 mice did not show histological abnormalities, whereas histological damage in the CA1 region of the hippocampus in SAMP8-control mice was observed. However, SAMP8-ACA mice were observed in a similar manner as SAMR1 mice. These findings confirm that ACA increases the serum concentrations of β-hydroxybutyric and levels and thus these fuels might contribute to the maintenance of the cognitive performance of SAMP8 mice.Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Keyword: energy

Comparison of diets enriched in stearic, oleic, and acids on inflammation, immune response, cardiometabolic risk factors, and fecal bile concentrations in mildly hypercholesterolemic postmenopausal women-randomized crossover trial.

Direct comparisons between SFAs varying in chain length, specifically (16:0) and stearic (18:0), relative to the latter\'s metabolic product, oleic (18:1), on cardiometabolic risk factors are limited.The aim of this study was to determine the relative comparability of diets enriched in , stearic , and oleic on inflammation and coagulation markers, T lymphocyte proliferation/ex-vivo cytokine secretion, plasma cardiometabolic risk factors, and fecal bile concentrations.Hypercholesterolemic postmenopausal women (n\xa0=\xa020, mean\xa0±\xa0SD age 64\xa0±\xa07 y, BMI 26.4\xa0±\xa03.4 kg/m2, LDL cholesterol\xa0≥\xa02.8 mmol/L) were provided with each of 3 diets [55% (%E) carbohydrate, 15%E protein, 30%E fat, with ∼50% fat contributed by , stearic , or oleic in each diet; 5 wk/diet phase] using a randomized crossover design with 2-wk washouts between phases. Outcome measures were assessed at the end of each phase.Fasting LDL-cholesterol and non-HDL-cholesterol concentrations were lower after the stearic and oleic diets than the diet (all P\xa0<\xa00.01). Fasting HDL-cholesterol concentrations were lower after the stearic diet than the and oleic diets (P\xa0<\xa00.01). The stearic diet resulted in lower lithocholic (P\xa0=\xa00.01) and total secondary bile (SBA) concentrations (P\xa0=\xa00.04) than the oleic diet. All other outcome measures were similar between diets. Lithocholic concentrations were positively correlated with fasting LDL-cholesterol concentrations (r\xa0=\xa00.33; P\xa0=\xa00.011). Total SBA, lithocholic , and deoxycholic concentrations were negatively correlated with fasting HDL cholesterol (r\xa0=\xa0-0.51 to -0.44; P\xa0<\xa00.01) concentrations and positively correlated with LDL cholesterol:HDL cholesterol (r =\xa00.37-0.54; P\xa0<\xa00.01) ratios.Dietary stearic and oleic had similar effects on fasting LDL-cholesterol and non-HDL-cholesterol concentrations and more favorable ones than . Unlike oleic , the hypocholesterolemic effect of stearic may be mediated by inhibition of intestinal hydrophobic SBA synthesis. These findings add to the data suggesting there should be a reassessment of current SFA dietary guidance and Nutrient Facts panel labeling.This trial was registered at clinicaltrials.gov as .Copyright © American Society for Nutrition 2019.

Keyword: energy

[CHANGES IN THE METABOLISM IN THE MYOCARDIUM OF RATS WITH ARTERIAL HYPERTENSION].

In the myocardium of the rats with arterial hypertension marked increase in the amount of unsaturated fatty acids and polyunsaturated fatty acids. Reducing the concentration of and increased levels of arachidonic is considered as one of the factors that lead to the development of deficit and oxidative stress. In rats, with hypertension myocardial lactate concentration increases in the cytoplasmic fraction and reducing the amount of ATP. The level in the cytoplasmic and mitochondrial fractions above benchmarks, indicating about the change of antioxidant systems of the body In the cytoplasm and mitochondria of cardiomyocytes of the rats with arterial hypertension marked decrease in the activity of antioxidant enzymes: NO-synthase, catalase, glutathione reductase, which causes metabolic changes of the myocardium.

Keyword: energy

Suitability of lemon balm (Melissa officinalis L.) extract rich in rosmarinic as a potential enhancer of functional properties in cupcakes.

Melissa officinalis (lemon balm) and its extracts have been frequently reported as possessing bioactive properties, offering the potential for use in development/enrichment of food products with additional functional capabilities, providing health benefits to consumers. The antioxidant, antibacterial and antifungal activity of lemon balm extract, as well as its potential hepatotoxicity were thoroughly evaluated. The extracts were then incorporated into cupcakes and their preserving effect, chemical composition, colour parameters and antioxidant activity were compared with those provided by potassium sorbate. In general, the variables with the largest differences among different storage times were level, sucrose, glucose, (C6:0) and oleic (C18:1n9). On the other hand, L (top), a (top), b (top), pH, capric (C10:0) and lauric (C12:0) showed the greatest variation according to cupcake formulation. The results observed indicate that the lemon balm extract rich in rosmarinic can provide advantageous functional properties to bakery products.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Growth, stool consistency and bone mineral content in healthy term infants fed sn-2-palmitate-enriched starter infant formula: A randomized, double-blind, multicentre clinical trial.

Palmitate in breast milk is predominantly located in the triacylglycerol sn-2 position, while infant formulae contain palmitate predominantly in the sn-1 and sn-3 positions. During digestion, palmitate in the sn-1 and sn-3 positions is hydrolyzed to free that can subsequently complex with calcium to form insoluble soaps; this may partially explain why formula-fed infants have harder stools than breast-fed infants.This large (n\xa0=\xa0488) randomized, double-blind, multicentre trial investigated whether increasing the sn-2 palmitate content of infant formula improves stool consistency and bone mineral content (measured by dual- x-ray absorptiometry), without affecting growth or health. From ∼1 week to 4 months of age, infants were exclusively fed one of three formulae: i) control formula (CF; 16% of total palmitate at sn-2; n\xa0=\xa0162), (ii) experimental formula 1 (EF1; 43% of total palmitate at sn-2; n\xa0=\xa0166) or (iii) experimental formula 2 (EF2; 51% of total palmitate at sn-2; n\xa0=\xa0160).Intention-to-treat analysis showed softer stools in both EF groups (vs. CF) at ages 2 weeks and 1 and 2 months (p\xa0≤\xa00.01), but not 3 and 4 months. At 4 months, all groups had similar growth outcomes while bone mineral content was significantly higher in EF1 (p\xa0=\xa00.0012) and EF2 (p\xa0=\xa00.0002) compared with CF. Comparison of reported adverse events up to 12 months revealed no differences among groups. All 3 infant formulae exhibited equally good digestive tolerance.Formulae enriched in sn-2 palmitate fed in early infancy are safe, improve stool consistency (from 2 weeks to 2 months) and increase bone mineral content (at 4 months).Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Keyword: energy

Encapsulation of soybean meal with fats enriched in and stearic acids: effects on rumen-undegraded protein and in vitro intestinal digestibility.

Fat coating of soybean meal (SBM) can reduce its protein degradability in the rumen, but the encapsulation of SBM with (PA) and stearic acids (SA) has not yet been investigated, despite both fatty acids are common sources in dairy cow diets. This study aimed to evaluate the effects of applying a novel method, using either 400 or 500\xa0g fat/kg (treatments FL40 and FL50, respectively), which was enriched in PA and SA at different ratios (100:0, 75:25, 50:50, 25:75 and 0:100), on physical and chemical characteristics, ruminal degradability, solubility and in vitro intestinal protein digestibility (IVIPD) of the obtained products. Encapsulation of SBM in fat resulted in greater mean particle size and lower bulk density and protein solubility than unprotected SBM (USBM). Treatment FL50 resulted in increased (p <\xa00.01) rumen-undegraded protein (RUP) compared to USBM. There were no differences in RUP of SBM when different PA: SA ratios were used. The mean RUP content of treatments FL40 and FL50 (306 and 349\xa0g/kg, respectively) was greater compared to USBM (262\xa0g/kg, p <\xa00.05), but lower than that for a standard heat-treated SBM (431\xa0g/kg). Values of IVIPD did not differ among SBM, heat-treated SBM and FL40 and FL50 samples, all being greater than 97.8%. In conclusion, encapsulation of SBM with fats enriched in PA and SA proved to be effective in reducing protein solubility and increasing RUP without depressing protein digestibility in the intestine. For validation of the method, in vivo research to investigate the effects of these products on the production of dairy cows is warranted.

Keyword: energy

Lipids characterization of ultrasound and microwave processed germinated sorghum.

Cereal crops and oilseeds provide diverse pool of fatty acids with characteristic properties. Sorghum (Sorghum bicolor (L.) Moench) provides the staple food with serving as main source of and protein. Germination of sorghum generally increases the nutritive value of seeds and the effects of germination on lipids composition of seeds vary greatly with processing conditions. Therefore, the current study was conducted to compare the effect of emerging processing techniques such as ultrasound (US) and microwave (MW) on fatty acids composition and oil yield of sorghum seeds before and after germination.Initially sorghum grains were soaked with 5% NaOCl (sodium hypochlorite) for surface sterilization. Afterwards, grains were soaked in excess water for 22\xa0h at room temperature and were divided into four portions. The first portion (100\xa0g grains) was subjected to germination without applying any microwave and ultrasonic treatment (T). Second portion was further divided into four groups (T, T, T, T) (100\xa0g of each group) and grains were subjected to ultrasonic treatments using two different ultrasonic intensities (US: 40%; US: 60%) within range of 0-100% and with two different time durations (t: 5\xa0min; t: 10\xa0min) at constant temperature. Third portion was also divided into four groups (T, T, T, T) (100\xa0g of each group) and exposed to microwave treatments at two different power levels (MW: 450 watt; MW: 700 watt) within the range of 100-900\xa0W for two different time durations (t: 15\xa0s; t: 30s). Similarly, fourth portion was divided into four groups (T, T, T, T) (100\xa0g of each group). Each group was exposed to both MW (MW, MW) (100-900 watt power) & US (US, US) (0-100% intensity) treatments at two different time levels (t, t). Then, germination was carried out and pre-treated raw and pre-treated germinated sorghum grains were analyzed for total oil yield, fatty composition and unsaturated fatty acids (Un-SFA)/saturated fatty acids (SFA) ratio by gas chromatography.The results revealed that oil yield in sorghum before and after germination ranged from 6.55 to 7.84% and 6.28 to 7.57%, respectively. All the microwave and ultrasound processed samples showed significant difference in oil yield than the raw sorghum grains. The highest tested yield was 7.84\xa0±\xa00.31% when combination of microwave power (700\xa0W) and ultrasound intensity (60%) was applied for 30s and 10\xa0min, respectively. The results further demonstrate that the raw sorghum contained (13.73\xa0±\xa00.10%), palmitoleic (0.43\xa0±\xa00.02%), stearic (1.07\xa0±\xa00.04%), oleic (37.15\xa0±\xa00.10%), linoleic (43.33\xa0±\xa00.21%), linolenic (1.55\xa0±\xa00.04%), arachidic (0.13\xa0±\xa00.01%) and eicosenoic (0.37\xa0±\xa00.02%), respectively. The highest fatty percentage for , stearic and arachidic was 13.75\xa0±\xa00.07%, 1.11\xa0±\xa00.09% and 0.15\xa0±\xa00.03% at 60% US intensity for 10\xa0min (T), respectively. Maximum amount observed was 1.60\xa0±\xa00.09% of linolenic while amount of eicosenoic decreased from 0.37\xa0±\xa00.02% to 0.31\xa0±\xa00.01% after processing. In case of applying combination of microwave and sonication treatments, the change in eicosenoic increased from 0.35\xa0±\xa00.02% to 0.40\xa0±\xa00.04% while there was no significant change in other fatty acids. The ungerminated sorghum oil possessed 14.93-15.05% and 82.83-83.12% of SFA and Un-SFA, respectively. After germination, percentage of saturated fatty acids increased (16.4-16.55%) while decreased for unsaturated fatty acids (80.13-80.56%) were noted.The results of the present study conclude that the yield of oil from sorghum grains increased by emerging processing. Fatty analysis of sorghum oil suggested that pre-treatment strategies will not affect the quality of the oil with respect to essential fatty acids content. Overall, the composition of saturated fatty in germinated grain is improved than ungerminated grains after processing.

Keyword: energy

Effects of high-fat diet and AMP-activated protein kinase modulation on the regulation of whole-body lipid metabolism.

Metabolic flexibility, the capacity to adapt to fuel availability for production, is crucial for maintaining whole-body homeostasis. An inability to adequately promote FA utilization is associated with lipid accumulation in peripheral tissues and contributes to the development of insulin resistance. In vivo assays to quantify whole-body lipid oxidation in mouse models of insulin resistance are lacking. We describe a method for assessing whole-body FA oxidation in vivo, as well as tissue-specific lipid uptake in conscious mice. The method relies on intravenous administration of [9,10-H(N)] combined with a non-β-oxidizable palmitate analog, [1-C]2-bromopalmitic . Pretreatment with etomoxir, a CPT1 inhibitor that prevents the shuttling of FAs into mitochondria, markedly reduced the appearance of the β-oxidation product HO in circulation and reduced lipid uptake by oxidative tissues including heart and soleus muscle. Whole-body fatty oxidation was unaltered between chow- or high-fat-fed WT and transgenic mice expressing a mutant form of the AMPK γ3 subunit (AMPKγ3) in skeletal muscle. High-fat feeding increased lipid oxidation in WT and AMPKγ3 transgenic mice. In conclusion, this technique allows for the assessment of the effect of pharmaceutical agents, as well as gene mutations, on whole-body FA oxidation in mice.Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Keyword: energy

Integrating Extracellular Flux Measurements and Genome-Scale Modeling Reveals Differences between Brown and White Adipocytes.

White adipocytes are specialized for storage, whereas brown adipocytes are specialized for expenditure. Explicating this difference can help identify therapeutic targets for obesity. A common tool to assess metabolic differences between such cells is the Seahorse Extracellular Flux (XF) Analyzer, which measures oxygen consumption and media acidification in the presence of different substrates and perturbagens. Here, we integrate the Analyzer\'s metabolic profile from human white and brown adipocytes with a genome-scale metabolic model to predict flux differences across the metabolic map. Predictions matched experimental data for the metabolite 4-aminobutyrate, the protein ABAT, and the fluxes for glucose, glutamine, and palmitate. We also uncovered a difference in how adipocytes dispose of nitrogenous waste, with brown adipocytes secreting less ammonia and more urea than white adipocytes. Thus, the method and software we developed allow for broader metabolic phenotyping and provide a distinct approach to uncovering metabolic differences.Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Role of fatty composites in the toxicity of titanium dioxide nanoparticles used in cosmetic products.

It has been recognized that the use of nanoparticles (NPs) in the cosmetic industry results in products with better efficacy and functionality. However, recent advances in molecular toxicology have revealed that NP exposure can promote cytotoxicity and oxidative damage, which has raised health concerns in the use of NPs in personal care products. Nevertheless, the mechanistic basis for the toxicity and safety of cosmetic NPs is poorly understood. The goal of the study was to determine the cytotoxicity and intracellular distribution of titanium dioxide (TiO2) NPs containing fatty composites (palmitoleic , , stearic and oleic ) commonly used in cosmetic products. Two types of cells, human fibroblast skin cells and adenocarcinoma lung cells, were exposed to either bare TiO2 NPs or TiO2 NPs mixed with fatty acids for up to 48 hr. NMR analysis confirmed that the fatty composites remained in the NPs after wash. The cytotoxicity of TiO2 NPs was determined by cell viability measurement using quantitative confocal microscopy, and the localization of two different forms of TiO2 NPs were assessed using electron spectroscopic imaging with transmission electron microscopy. TiO2 NPs containing fatty acids posed significantly reduced cytotoxicity (80-88% decreases) than bare NPs in both cell types. Furthermore, there was less intracellular penetration of the NPs containing fatty composites compared with bare NPs. These results provide important insights into the role of fatty acids in protecting the cells from possible toxicity caused by NPs used in the production of cosmetic products.

Keyword: energy

Stabilizing mixed fatty and phthalate ester monolayer on artificial seawater.

Phthalate esters which are widely used as industrial chemicals have become widespread contaminants in the marine environment. However, little information is available on the interfacial behavior of phthalate esters in the seawater, where contaminants generally occur at elevated concentrations and have the potential to transfer into the atmosphere through wave breaking on sea surface. We used artificial seawater coated with fatty acids to simulate sea surface microlayer in a Langmuir trough. The interactions of saturated fatty acids (stearic (SA) and (PA)) with one of the most abundant phthalate esters (di-(2-ethylhexyl) phthalate (DEHP)), were investigated under artificial seawater and pure water conditions. Pure DEHP monolayer was not stable, while more stable mixed monolayers were formed by SA and DEHP on the artificial seawater at relatively low surface pressure. Sea salts in the subphase can lower the excess Gibbs free to form more stable mixed monolayer. Among the ten components in the sea salts, Ca ions played the major role in condensation of mixed monolayer. The condensed characteristic of the mixed SA (or PA)/DEHP monolayers suggested that the hydrocarbon chains were ordered on artificial seawater. By means of infrared reflection-absorption spectroscopy (IRRAS), we found that multiple sea salt mixtures induced deprotonated forms of fatty acids at the air-water interface. Sea salts can improve the stability and lifetime of mixed fatty and phthalate ester monolayer on aqueous droplets in the atmosphere. Interfacial properties of mixed fatty and phthalate ester monolayers at the air-ocean interface are important to help understand their behavior and fate in the marine environment.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: energy

Enhanced Fatty Scavenging and Glycerophospholipid Metabolism Accompany Melanocyte Neoplasia Progression in Zebrafish.

Alterations in lipid metabolism in cancer cells impact cell structure, signaling, and metabolism, making lipid metabolism a potential diagnostic marker and therapeutic target. In this study, we combined PET, desorption electrospray ionization-mass spectrometry (DESI-MS), nonimaging MS, and transcriptomic analyses to interrogate changes in lipid metabolism in a transgenic zebrafish model of oncogenic RAS-driven melanocyte neoplasia progression. Exogenous fatty uptake was detected in melanoma tumor nodules by PET using the surrogate tracer 14(R,S)-18F-fluoro-6-thia-heptadecanoic ([18F]-FTHA), consistent with upregulation of genes associated with fatty uptake found through microarray analysis. DESI-MS imaging revealed that FTHA uptake in tumors was heterogeneous. Transcriptome and lipidome analyses further highlighted dysregulation of glycerophospholipid pathways in melanoma tumor nodules, including increased abundance of phosphatidyl ethanolamine and phosphatidyl choline species, corroborated by DESI-MS, which again revealed heterogeneous phospholipid composition in tumors. Overexpression of the gene encoding lipoprotein lipase (LPL), which was upregulated in zebrafish melanocyte tumor nodules and expressed in the majority of human melanomas, accelerated progression of oncogenic RAS-driven melanocyte neoplasia in zebrafish. Depletion or antagonism of LPL suppressed human melanoma cell growth; this required simultaneous fatty synthase (FASN) inhibition when FASN expression was also elevated. Collectively, our findings implicate fatty acquisition as a possible therapeutic target in melanoma, and the methods we developed for monitoring fatty uptake have potential for diagnosis, patient stratification, and monitoring pharmacologic response. SIGNIFICANCE: These findings demonstrate the translational potential of monitoring fatty uptake and identify lipoprotein lipase as a potential therapeutic target in melanoma.©2019 American Association for Cancer Research.

Keyword: energy

Utilization of lactic in human myotubes and interplay with glucose and fatty metabolism.

Once assumed only to be a waste product of anaerobe glycolytic activity, lactate is now recognized as an source in skeletal muscles. While lactate metabolism has been extensively studied in vivo, underlying cellular processes are poorly described. This study aimed to examine lactate metabolism in cultured human myotubes and to investigate effects of lactate exposure on metabolism of oleic and glucose. Lactic , fatty and glucose metabolism were studied in myotubes using [C(U)]lactic , [C]oleic and [C(U)]glucose, respectively. Myotubes expressed both the MCT1, MCT2, MCT3 and MCT4 lactate transporters, and lactic was found to be a substrate for both glycogen synthesis and lipid storage. Pyruvate and inhibited lactic oxidation, whilst glucose and α-cyano-4-hydroxycinnamic inhibited lactic uptake. Acute addition of lactic inhibited glucose and oleic oxidation, whereas oleic uptake was increased. Pretreatment with lactic for 24\u2009h did not affect glucose or oleic metabolism. By replacing glucose with lactic during the whole culturing period, glucose uptake and oxidation were increased by 2.8-fold and 3-fold, respectively, and oleic oxidation was increased 1.4-fold. Thus, lactic has an important role in metabolism of human myotubes.

Keyword: energy

Diosgenin ameliorates -induced lipid accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is characterized by excessive hepatic lipid accumulation. Many studies have suggested that lipid overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on metabolism, but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic fatty liver. The effects of DSG on lipid metabolism, glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, lipid consumption and lipid synthesis in LO2 cells was investigated.The results indicated that induced obvious lipid accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular lipid content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of lipid synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and fatty synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular lipid accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing fatty β-oxidation and decreasing lipid synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: energy

Homology modeling and docking studies of a Δ9-fatty desaturase from a Cold-tolerant sp. AMS8.

Membrane-bound fatty desaturases perform oxygenated desaturation reactions to insert double bonds within fatty acyl chains in regioselective and stereoselective manners. The Δ9-fatty desaturase strictly creates the first double bond between C9 and 10 positions of most saturated substrates. As the three-dimensional structures of the bacterial membrane fatty desaturases are not available, relevant information about the enzymes are derived from their amino sequences, site-directed mutagenesis and domain swapping in similar membrane-bound desaturases. The cold-tolerant sp. AMS8 was found to produce high amount of monounsaturated fatty acids at low temperature. Subsequently, an active Δ9-fatty desaturase was isolated and functionally expressed in . In this paper we report homology modeling and docking studies of a Δ9-fatty desaturase from a Cold-tolerant sp. AMS8 for the first time to the best of our knowledge. Three dimensional structure of the enzyme was built using MODELLER version 9.18 using a suitable template. The protein model contained the three conserved-histidine residues typical for all membrane-bound desaturase catalytic activity. The structure was subjected to minimization and checked for correctness using Ramachandran plots and ERRAT, which showed a good quality model of 91.6 and 65.0%, respectively. The protein model was used to preform MD simulation and docking of using CHARMM36 force field in GROMACS Version 5 and Autodock tool Version 4.2, respectively. The docking simulation with the lowest binding , -6.8 kcal/mol had a number of residues in close contact with the docked namely, Ile26, Tyr95, Val179, Gly180, Pro64, Glu203, His34, His206, His71, Arg182, Thr85, Lys98 and His177. Interestingly, among the binding residues are His34, His71 and His206 from the first, second, and third conserved histidine motif, respectively, which constitute the active site of the enzyme. The results obtained are in compliance with the activity of the Δ9-fatty desaturase on the membrane phospholipids.

Keyword: energy

AMPK regulates metabolism through the SIRT1 signaling pathway to improve myocardial hypertrophy.

We investigated the correlations of adenosine monophosphate-activated protein kinase (AMPK), Silence information regulator 1 (SIRT1) and metabolism with myocardial hypertrophy.Myocardial hypertrophy experimental model was established via transverse aortic constriction (TAC)-induced myocardial hypertrophy and phenylephrine (PE)-induced hypertrophic myocardial cell culture. After activation of AMPK, the messenger ribonucleic (mRNA) expressions in myocardial tissue- and myocardial cell hypertrophy-related genes, atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MHC), were detected. The production rate of 14C-labeled 14CO2 from was quantitatively determined to detect the fatty and glucose oxidation of hypertrophic myocardial tissues or cells, and the glucose uptake of myocardial cells was studied using [14C] glucose. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to detect the changes in SIRT1 mRNA and protein expressions in hypertrophic myocardial tissues. Moreover, SIRT1 small interfering ribonucleic (siRNA) was used to interfere in SIRT1 expression to further investigate the role of SIRT1 in the effect of AMPK activation on myocardial hypertrophy.AMPK activation could significantly reduce the mRNA expressions of ANP and β-MHC in vitro and in vivo. AMPK could increase the ejection fraction (EF) and decrease the protein synthesis rate in myocardial cells in mice with myocardial hypertrophy. Besides, AMPK activation could increase the fatty oxidation, improve the glucose uptake and reduce the glucose oxidation. After AMPK activation, both SIRT1 mRNA and protein expressions in hypertrophic myocardial tissues and myocardial cells were increased. After SIRT1 siRNA was further used to interfere in SIRT1 expression in myocardial cells, it was found that mRNA expressions and protein synthesis rates of ANP and β-MHC were increased.The activation of AMPK can inhibit the myocardial hypertrophy, which may be realized through regulating the myocardial metabolism via SIRT1 signaling pathway.

Keyword: energy

and nitrogen partitioning in dairy cows at low or high metabolizable protein levels is affected differently by postrumen glucogenic and lipogenic substrates.

This study tested the effects of from glucogenic (glucose; GG) or lipogenic (palm olein; LG) substrates at low (LMP) and high (HMP) metabolizable protein levels on whole-body and N partitioning of dairy cattle. Six rumen-fistulated, second-lactation Holstein-Friesian dairy cows (97 ± 13 d in milk) were randomly assigned to a 6 × 6 Latin square design in which each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of rest. A total mixed ration consisting of 42% corn silage, 31% grass silage, and 27% concentrate (dry matter basis) was formulated to meet 100 and 83% of net and metabolizable protein requirements, respectively, and was fed at 90% of ad libitum intake by individual cow. Abomasal infusion treatments were saline (LMP-C), isoenergetic infusions (digestible basis) of 1,319 g/d of glucose (LMP-GG), 676 g/d of palm olein (LMP-LG; major fatty constituents are , oleic, and linoleic ), or 844 g/d of essential AA (HMP-C), or isoenergetic infusions of 1,319 g/d of glucose + 844 g/d of essential AA (HMP-GG) or 676 g/d of palm olein + 844 g/d of essential AA (HMP-LG). The experiment was conducted in climate respiration chambers to determine and N balance in conjunction with milk production and composition, nutrient digestibility, and plasma constituents. Infusion of GG and LG decreased dry matter intake, but total gross intake from the diet plus infusions was not affected by GG or LG. Furthermore, GG or LG did not affect total milk, protein, or lactose yields. Infusing GG or LG at the HMP level did not affect milk production differently than at the LMP level. Infusion of GG stimulated retention in body tissue, increased plasma glucose and insulin concentrations, decreased lipogenic metabolites in plasma, and decreased milk fat yield and milk output. Nitrogen intake decreased and milk N efficiency increased in response to GG, and N retention was not affected. Infusion of LG tended to increase metabolizable intake, increased milk fat yield and milk output, increased plasma triacylglycerides and long-chain fatty concentrations, and had no effect on retention. Infusion of LG decreased N intake but did not affect milk N efficiency or N retention. Compared with the LMP level, the HMP level increased dry matter intake, gross and metabolizable intake, and total milk, fat, protein, and lactose yields. Milk output increased at the HMP level, and protein level did not affect total retention. Heat production increased at the HMP level, but only when GG and LG were infused. The HMP level increased N intake, milk N output, and plasma urea concentration, tended to increase N retention, and decreased milk N efficiency. Regardless of protein level, GG promoted retention and improved milk N efficiency, but not through increased milk protein yield. Infusion of LG partitioned extra intake into milk and had no effect on milk N efficiency.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

(L.) L. subsp. .: Nutritional Characterization.

The habit of eating wild plants in Europe is often associated with times of famine; an example of such is the nectar of (L.) L., a parasitic plant. To the authors\' best knowledge, there are no studies on its nutritional and chemical composition; thus, the whole (L.) L. subsp. Wettst. plant (CH) and its nectar (NCH) were nutritionally and chemically characterized. The proximate composition of CH and NCH were very similar in terms of , ash, and carbohydrate content. Protein and fat were approximately 2-fold higher in NCH, and crude fiber was 4.6-fold higher in CH compared to NCH. Fructose, glucose, sucrose, and trehalose were the free sugars present in both samples. Oxalic, malic, and citric acids were the identified organic acids in both samples, with citric as the most abundant molecule. For both samples, polyunsaturated and saturated fatty acids (PUFA and SFA, respectively) predominate over monounsaturated fatty acids (MUFA) due to the significant contribution of linoleic and acids, respectively. However, unsaturated fatty acids (UFA) prevail over SFA in CH and NCH. Therefore, CH proved to be an excellent source of nutritional compounds, which supports its use during past periods of scarcity.

Keyword: energy

High-fat diet feeding and increase CRC growth in β2AR-dependent manner.

Epidemiology studies indicate that consumption of high-fat diet (HFD) is directly associated with the development of colorectal cancer (CRC). However, the exact component in HFD and the mechanism underlying its effect on CRC growth remained unclear. Our study shows that HFD feeding increases β2AR expression in the xenograft tissues of CRC-bearing mouse model; the elevated β2AR expression is reduced when HFD is replaced by control diet, which strongly suggests an association between HFD feeding and β2AR expression in CRC. HFD feeding increases and stearic levels in CRC; however, only increases β2AR expression, which is dependent upon Sp1. β2AR plays the dominant role in promoting CRC cell proliferation among all the β-AR subtypes. More importantly, knockout of β2AR or knockdown of Sp1 abolishes the increased CRC cell proliferation, suggesting increases CRC cell proliferation in β2AR-dependent manner. HFD or -rich diet (PAD) also fails to increase the tumor growth in xenograft mouse models bearing β2AR-knockout CRC cells. β2AR promotes CRC growth by increasing the phosphorylation of HSL at the residue S552. The phosphorylated and activated HSL (S552) changes the metabolic phenotype of CRC and increases production, which promotes CRC growth. Our study has revealed the unique tumorigenic properties of in promoting CRC growth, and have delineated the underlying mechanism of action. We are also the first to report the linkage between HFD feeding and β-adrenergic signaling pathway in relation to CRC growth.

Keyword: energy

Fatty composition in serum cholesterol esters and phospholipids is linked to visceral and subcutaneous adipose tissue content in elderly individuals: a cross-sectional study.

Visceral adipose tissue (VAT) and truncal fat predict cardiometabolic disease. Intervention trials suggest that saturated fatty acids (SFA), e.g. , promote abdominal and liver fat storage whereas polyunsaturated fatty acids (PUFA), e.g. linoleic , prevent fat accumulation. Such findings require investigation in population-based studies of older individuals. We aimed to investigate the relationships of serum biomarkers of PUFA intake as well as serum levels of , with abdominal and total adipose tissue content.In a population-based sample of 287 elderly subjects in the PIVUS cohort, we assessed fatty composition in serum cholesterol esters (CE) and phospholipids (PL) by gas chromatography and the amount of VAT and abdominal subcutaneous (SAT) adipose tissue by magnetic resonance imaging (MRI), liver fat by MR spectroscopy (MRS), and total body fat, trunk fat and leg fat by dual- X-ray absorptiometry (DXA). Insulin resistance was estimated by HOMA-IR.VAT and trunk fat showed the strongest correlation with insulin resistance (r\u2009=\u20090.49, P\u2009<\u20090.001). Linoleic in both CE and PL was inversely related to all body fat depots (r\u2009=\u2009-0.24 to -0.33, P\u2009<\u20090.001) including liver fat measured in a sub-group (r\u2009=\u2009-0.26, P\u2009<\u20090.05, n\u2009=\u200973), whereas n-3 PUFA showed weak inverse (18:3n-3) or positive (20:5n-3) associations. in CE, but not in PL, was directly correlated with VAT (r\u2009=\u20090.19, P\u2009<\u20090.001) and trunk fat (r\u2009=\u20090.18, P\u2009=\u20090.003). Overall, the significant associations remained after adjusting for intake, height, alcohol, sex, smoking, education and physical activity. The inverse correlation between linoleic and VAT remained significant after further adjustment for total body fat.Serum linoleic is inversely related to body fat storage including VAT and trunk fat whereas was less consistently but directly associated, in line with recent feeding studies. Considering the close link between VAT and insulin resistance, a potential preventive role of plant-based PUFA in VAT accumulation warrants further study.

Keyword: energy

Apoptotic germ cells regulate Sertoli cell lipid storage and fatty oxidation

The presence of lipid droplets (LDs) and the utilization of fatty acids (FAs) as a source of are Sertoli cell (SC) putative characteristics. It is well known that SCs can phagocyte and degrade apoptotic germ cells (AGCs) resulting in increasing lipid content and ATP levels. A relationship between the regulation of lipid storage and of lipid oxidation in SC might be envisaged. The aim of this study was to analyze whether AGCs and FAs are able to simultaneously regulate molecular mechanisms involved in lipid storage and in FA oxidation in SC. The experimental model utilized in this study consisted in SC cultures obtained from 20-day-old rats that were co-cultured with AGC or treated with (PA, 500 μM) for 24 and 48 h. AGC and PA increase LD, triacylglycerol (TAG) content and mRNA levels of Plin1, Plin2, Plin3 (proteins involved in TAG storage). Simultaneously, AGC and PA rise the extent of FA oxidation and mRNA levels of Cpt1 and Lcad (proteins involved in FA degradation). Results also show that peroxisome proliferator-activated receptor (PPAR) transcriptional activity, transcription factor which participate in lipid metabolism regulation, increases by AGC and PA treatment in SC. Additionally, the presence of a PPARg antagonist decreases the upregulation of LD content and Plin1 expression. Similarly, the presence of a PPARb/d antagonist reduces the increase in FA oxidation and Cpt1 mRNA levels. Altogether these results suggest that AGC and FA, which probably generate PPAR ligands, regulate lipid storage and fatty utilization, contributing to the homeostasis in the seminiferous tubules.

Keyword: energy

Calculation of apparent pK values of saturated fatty acids with different lengths in DOPC phospholipid bilayers.

We performed all-atom molecular dynamics simulations and calculated free profiles and apparent pKa values for neutral and anionic forms of single myristic (C14:0), (C16:0) and stearic (C18:0) fatty embedded in a DOPC bilayer and explicit water solvent. We showed that the neutral forms of the fatty acids are stabilized inside the bilayer by hydrogen bonding of a fatty carboxylic group with DOPC phosphate and carbonyl groups. In contrast to the neutral form, the anionic forms of the fatty acids are shifted towards the water-membrane interface and are instead stabilized by hydrogen bonding to interfacial water. By using umbrella sampling simulations, we calculated free energies of stabilization and revealed that the free of stabilization inside the bilayer increases with the chain length for both the neutral and deprotonated forms. On the other hand, the free energies of flip-flop of both the neutral and anionic forms are constant upon the prolongation of the fatty . Based on the free curves, we also calculated apparent fatty pKa,app values in the bilayer, which are 7.0, 7.2 and 6.3 for myristic, and stearic and are increased by several pKa units compared to the corresponding pKa values in water. By further analysis of the calculated curves we found that spontaneous protonation of fatty anions takes place in the bilayer interior at ca. 1.4 nm from the bilayer center for all studied fatty acids.

Keyword: energy

Selective synthesis of human milk fat-style structured triglycerides from microalgal oil in a microfluidic reactor packed with immobilized lipase.

Human milk fat-style structured triacylglycerols were produced from microalgal oil in a continuous microfluidic reactor packed with immobilized lipase for the first time. A remarkably high conversion efficiency was demonstrated in the microreactor with reaction time being reduced by 8 times, Michaelis constant decreased 10 times, the lipase reuse times increased 2.25-fold compared to those in a batch reactor. In addition, the content of at sn-2 position (89.0%) and polyunsaturated fatty acids at sn-1, 3 positions (81.3%) are slightly improved compared to the product in a batch reactor. The increase of melting points (1.7°C) and decrease of crystallizing point (3°C) implied higher quality product was produced using the microfluidic technology. The main cost can be reduced from $212.3 to $14.6 per batch with the microreactor. Overall, the microfluidic bioconversion technology is promising for modified functional lipids production allowing for cost-effective approach to produce high-value microalgal coproducts.Copyright © 2016 Elsevier Ltd. All rights reserved.

Keyword: energy

Compound C attenuates NLRP3 inflammasome despite AMPK knockdown in LPS plus palmitate-induced THP-1 cells.

NLRP3 inflammasome is a key contributor to obesity-related insulin resistance and type 2 diabetes (T2D). Adenosine monophosphate-activated protein kinase (AMPK) is a principle intracellular sensor exerting protective effect against T2D. Strikingly, compound C, an inhibitor of AMPK, considerably inhibited the secretion of IL-1β when THP-1 cells were stimulated with LPS plus (PA). The underlying mechanism was examined with respect to the effect of compound C on NLRP3 inflammasome, a multiprotein complex which controls the processing and production of IL-1β. Interestingly, compound C significantly attenuated the activation of NLRP3 inflammasome. This phenomenon was reproduced in AMPK siRNA-transfected THP-1 cells, indicating that compound C exerts this function despite AMPK knockdown. Also, it significantly suppresses the mitochondria-generated reactive oxygen species (ROS) required for NLRP3 inflammasome activation. In conclusion, compound C was shown to significantly attenuate the NLRP3 inflammasome despite AMPK knockdown, rendering it as the novel target of compound C. Potentially, compound C attenuates NLRP3 inflammasome through the suppression of mitochondrial ROS production. These findings offer initial evidence into compound C as a novel pharmacological agent with significant therapeutic potential in NLRP3 inflammasome-related disorders, including obesity, insulin resistance, and T2D. Thus, further studies are essential to identify the effect of compound C on these diseases in vitro.

Keyword: energy

Loss of biliverdin reductase-A promotes lipid accumulation and lipotoxicity in mouse proximal tubule cells.

Obesity and increased lipid availability have been implicated in the development and progression of chronic kidney disease. One of the major sites of renal lipid accumulation is in the proximal tubule cells of the kidney, suggesting that these cells may be susceptible to lipotoxicity. We previously demonstrated that loss of hepatic biliverdin reductase A (BVRA) causes fat accumulation in livers of mice on a high-fat diet. To determine the role of BVRA in mouse proximal tubule cells, we generated a CRISPR targeting BVRA for a knockout in mouse proximal tubule cells (BVRA KO). The BVRA KO cells had significantly less metabolic potential and mitochondrial respiration, which was exacerbated by treatment with , a saturated fatty . The BVRA KO cells also showed increased intracellular triglycerides which were associated with higher fatty uptake gene cluster of differentiation 36 as well as increased de novo lipogenesis as measured by higher neutral lipids. Additionally, neutrophil gelatinase-associated lipocalin 1 expression, annexin-V FITC staining, and lactate dehydrogenase assays all demonstrated that BVRA KO cells are more sensitive to -induced lipotoxicity than wild-type cells. Phosphorylation of BAD which plays a role in cell survival pathways, was significantly reduced in -treated BVRA KO cells. These data demonstrate the protective role of BVRA in proximal tubule cells against saturated fatty -induced lipotoxicity and suggest that activating BVRA could provide a benefit in protecting from obesity-induced kidney injury.

Keyword: energy

Association of dietary factors with severity of coronary artery disease.

Cardiovascular disease has taken epidemic proportions during past decades. Cardiovascular risk factors contribute to progression of coronary lesions, worsening the patient\'s prognosis. This study was planned to analyze the association of dietary factors with severity of coronary artery disease (CAD) in Indian patients.Three hundred patients with known coronary disease above the age of 25 years were included in this study. Blood samples were collected for biochemical markers. Patients were stratified according to severity of CAD [number of vessel involved-single (SVD), double (DVD), triple (TVD)].Mean age of the patient was 60.9\xa0±\xa012.4 years. Subjects with TVD, DVD, SVD in the study were 52.3%, 25.3% and 22.3% respectively. Patients with TVD had higher body mass index, triglycerides, HOMA-Insulin Resistance, hsCRP and lower high density cholesterol. Diabetes mellitus, hypertension and dyslipidemia were more common in TVD patients. Among macronutrients, patients with TVD had higher intake of carbohydrate and lower intake of protein and dietary fibers. There was no association of total fat intake with CAD, however, intake of was higher among patients with TVD. Intake of vitamins namely niacin, riboflavin, thiamine, B6, and vitamin-C decreased with increase in severity. With increase in severity of CAD, mineral intake (potassium, calcium, magnesium, phosphorus, sulfur, iron, chromium, copper, manganese, and zinc) decreased.Dietary factors are associated with severity of coronary artery disease. Low intake of protein, fiber, vitamins, minerals and high intake of carbohydrate and fat was associated with higher probability of having severe CAD.Copyright © 2016 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved.

Keyword: energy

Valorization of sewage sludge via non-catalytic transesterification.

To seek a way to valorize sewage sludge (SS), it was chosen as a raw material for biodiesel production. As such, non-catalytic transesterification of dried SS was carried out, to enhance its value. Note that picking a waste material such as SS as an inexpensive lipid feedstock for biodiesel production, without lipid extraction, greatly increases the economic viability of biodiesel. Also, to enhance biodiesel sustainability, ethanol (EtOH) was employed as the acyl acceptor, in this study, and this was experimentally justified by results showing that employing EtOH as an acyl acceptor provided an effective means for compensating for the lower heating value arising from the large amount of (C) in SS. This study experimentally proved that the fatty ethyl ester (FAEE) yield at 380\u202f°C reached up to 13.33\u202fwt% of dried SS. Given that the lipid content of dried SS is 14.01\u202f±\u202f0.64\u202fwt%, the FAEE yield of 13.33\u202fwt% implied that 95.14\u202fwt% of lipid in dried SS had been converted into FAEEs. The introduced SS valorization in this study offered an excellent opportunity to address diverse environmental hazards arising from SS and associated emerging contaminants. Given that the optimal temperature for the non-catalytic conversion for biodiesel production from SS was found to be 380\u202f°C, emerging contaminants, such as microplastics and antimicrobials, were simultaneously degraded, due to their inferior thermal stabilities. Lastly, considering that the introduced biodiesel conversion process is thermally induced, the SS reside after the biodiesel conversion process can be further used in thermo-chemical processes as raw materials for gasification and pyrolysis (future work).Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.

Keyword: energy

Gene expressions and metabolomic research on the effects of polyphenols from the involucres of Castanea mollissima Blume on heat-stressed broilers chicks.

To study the effects of polyphenolic extract from involucres of Castanea mollissima Blume ( PICB: ), a novel approach using gene expression by real time polymerase chain reaction ( REAL-TIME PCR: ) coupled with metabolomic profiling technique was established to explain the mechanism of PICB on heat-stressed broiler chicks. Four thousand 28-day-old male Arbor Acres (AA) broilers were randomly assigned to 5 groups (4 replicates / group, 20 chicks / replicate), in which group 1 was normal control group fed with basic ration; groups 2, 3, 4, and 5 were fed with the basic ration with a supplementation of 0.2% Vitamin C ( VC: ), or 0.2%, 0.3%, or 0.4% of PICB respectively. After 1 wk of adaptation, heat stress was applied for 7 consecutive days. On d 3 and d 7 of heat stress, the chicks were sacrificed and sampled. The mRNA expression of heat stress protein 70 (HSP70), glutathione peroxidase ( GSH-PX: ), ornithine decarboxylase ( ODC: ), epidermal growth factor ( EGF: ) and epidermal growth factor receptor ( EGFR: ) were detected by real-time PCR using samples from jejunum mucosa. The serum and jejunum mucosa metabolomic profiles of PICB group showing best antioxidative effects and control group at d 3 were studied using the method of the gas chromatography - time of flight mass spectrometry ( GT-TOF-MS: ), followed by principal component analysis and partial least squares-discriminate analysis. Potential biomarkers were found using Student\'s t-test. The results showed mRNA expressions of HSP70, GSH-Px, ODC, EGF, and EGFR were altered by the supplementation of PICB. PICB exhibited antioxidative and growth promoting effects, and 0.3% PICB supplementation level exhibited the best. Three metabolites in the serum and 5 in the jejunum mucosa were identified as potential biomarkers. They were considered to be in accordance with antioxidative and growth promoting effects of PICB, which involved in the metabolism (sorbitol, ), carbohydrate metabolism, amino acids metabolism (serine, L-ornithine), glutathione metabolism (glutamate, L-ornithine), GnRH signaling pathway (inositol), etc. These findings provided novel insights into our understanding of molecular mechanism of PICB effects on heat-stressed chicks.© 2016 Poultry Science Association Inc.

Keyword: energy

DGAT1-Dependent Lipid Droplet Biogenesis Protects Mitochondrial Function during Starvation-Induced Autophagy.

Lipid droplets (LDs) provide an "on-demand" source of fatty acids (FAs) that can be mobilized in response to fluctuations in nutrient abundance. Surprisingly, the amount of LDs increases during prolonged periods of nutrient deprivation. Why cells store FAs in LDs during an crisis is unknown. Our data demonstrate that mTORC1-regulated autophagy is necessary and sufficient for starvation-induced LD biogenesis. The ER-resident diacylglycerol acyltransferase 1 (DGAT1) selectively channels autophagy-liberated FAs into new, clustered LDs that are in close proximity to mitochondria and are lipolytically degraded. However, LDs are not required for FA delivery to mitochondria but instead function to prevent acylcarnitine accumulation and lipotoxic dysregulation of mitochondria. Our data support a model in which LDs provide a lipid buffering system that sequesters FAs released during the autophagic degradation of membranous organelles, reducing lipotoxicity. These findings reveal an unrecognized aspect of the cellular adaptive response to starvation, mediated by LDs.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: energy

Effects of timing of supplementation during early lactation on nutrient digestibility, balance, and metabolism of dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation during early lactation on nutrient digestibility, intake and balance, and metabolic responses of dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh (FR) period (1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a C16:0-supplemented diet [PA; 1.5% of diet dry matter (DM)]. During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA diet (1.5% of diet DM) in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, compared with CON, PA increased DM digestibility by 3.0 percentage units and neutral detergent fiber (NDF) digestibility by 4.4 percentage units, and the increase in these variables was consistent over time. Although PA did not affect 18-carbon fatty (FA) digestibility, it decreased 16-carbon FA digestibility by 10.8 percentage units and total FA digestibility by 4.7 percentage units compared with CON. We observed a tendency for an interaction between treatment and time for total FA digestibility and 16-carbon FA digestibility due to the difference in FA digestibility between PA and CON reducing over time. Compared with CON, PA increased digestible intake by 3.9 Mcal/d, metabolizable intake by 3.5 Mcal/d, and net for lactation intake by 2.5 Mcal/d. The PA diet also increased milk output, negative balance, and plasma nonesterified fatty concentration and reduced plasma insulin concentration. We also observed a tendency for an interaction between treatment and time for balance due to cows receiving the PA treatment being in a greater negative balance over time. During the PK period, PA increased DM digestibility by 2.9 percentage units and NDF digestibility by 3.5 percentage units compared with CON. Although PA decreased 16-carbon FA digestibility by 7.0 percentage units, PA did not affect 18-carbon FA digestibility or total FA digestibility. Feeding PA during the PK period increased intake and milk output and did not affect balance. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased DM and NDF digestibilities and intake compared with a control diet containing no supplemental fat. Feeding C16:0 markedly increased milk output in both the FR and PK periods but increased negative balance only in the FR period.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Metabolic transition of milk triacylglycerol synthesis in response to varying levels of palmitate in porcine mammary epithelial cells.

Milk in mammals is a key source of lipids for offspring, providing both critical and essential fatty acids. For lactating sows, is one of the most abundant fatty acids in milk, providing 10~12% of the suckling pig total dietary supply. However, the effects of exogenous on milk fat synthesis in sow mammary glands are not well-known. In this study, we investigated the effects of on lipogenic genes in porcine mammary epithelial cells (pMECs) to explore the role of exogenous in mediating milk triacylglycerols (TAG) synthesis.Porcine mammary epithelial cells were cultured for 24\xa0h in the presence of different concentrations of palmitate (0, 25, 50, 100, 200, 400, and 600\xa0μM). The effect of palmitate on cell viability was tested via MTT assay. Intracellular lipid accumulation was measured through Oil Red O staining, and TAG levels were quantified by enzymatic colorimetric methods. Expression of genes and proteins involved in milk fat biosynthesis were assayed with quantitative real-time polymerase chain reaction (PCR) and Western blotting, respectively.Incubation with palmitate promoted cellular lipid synthesis in a dose-dependent manner, as reflected by the increased TAG content and enhanced formation of cytosolic lipid droplets. The increased lipid synthesis by palmitate was probably attributable to the upregulated mRNA expression of genes associated with milk fat biosynthesis, including long-chain fatty uptake (, ), intracellular activation and transport , ), TAG synthesis (, , ), lipid droplet formation (), and regulation of transcription (). Western blot analysis of CD36 and DGAT1 proteins confirmed the increased lipid synthesis with increasing incubation of palmitate. However, the genes involved in fatty de novo synthesis (, ), fatty desaturation (), and regulation of transcription (, ) were inversely affected by incubation with increasing concentrations of palmitate. Western blot analysis of ACACA protein confirmed this decrease associated with increasing levels of palmitate.Results from this study suggest that palmitate stimulated the cytosolic TAG accumulation in pMECs, probably by promoting lipogenic genes and proteins that are involved in lipid synthesis. However, addition of palmitate decreased the fatty de novo synthesis in pMECs.

Keyword: energy

Effects of timing of supplementation on production responses of early-lactation dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation on production responses of early-lactation dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh period (FR; 1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a diet supplemented with C16:0 (, PA; 1.5% of diet dry matter). During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA (1.5% of diet dry matter) diet in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, we did not observe treatment differences for dry matter intake or milk yield. Compared with CON, PA increased the yield of 3.5% fat-corrected milk by 5.30 kg/d, yield of -corrected milk (ECM) by 4.70 kg/d, milk fat content by 0.41% units, milk fat yield by 280 g/d, and protein yield by 100 g/d. The increase in milk fat associated with the PA treatment during the FR period occurred due to an increase in yield of 16-carbon milk fatty acids (FA) by 147 g/d (derived from both de novo synthesis and extraction from plasma) and preformed milk FA by 96 g/d. Compared with CON, PA decreased body weight (BW) by 21 kg and body condition score (BCS) by 0.09 units and tended to increase BW loss by 0.76 kg/d. Although PA consistently increased milk fat yield and ECM over time, a treatment × time interaction was observed for BW and BCS due to PA inducing a greater decrease in BW and BCS after the second week of treatments. Feeding PA during the PK period increased milk yield by 3.45 kg/d, yield of 3.5% fat-corrected milk by 4.50 kg/d, yield of ECM by 4.60 kg/d, milk fat content by 0.22% units, milk fat yield by 210 g/d, protein yield by 140 g/d, and lactose yield by 100 g/d but tended to reduce BW by 10 kg compared with CON. Also, during the PK period we observed an interaction between diet fed in the FR and PK periods for milk fat yield due to feeding PA during the PK period increasing milk fat yield to a greater extent in cows that received the CON diet (+240 g/d) rather than the PA diet (+180 g/d) during the FR period. This difference was associated with the yield of preformed FA because feeding PA during the PK period increased the yield of preformed milk FA only in cows that received the CON diet during the FR period. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased the yield of ECM in both the FR and PK periods compared with a control diet. For some variables, the effect of feeding C16:0 was affected by timing of supplementation because milk yield increased only during the PK period and BW decreased to a greater extent in the FR period. Regardless of diet fed in the FR period, feeding a C16:0 supplement during the PK period increased yields of milk and milk components.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: energy

Autophagy activation by Jiang Zhi Granule protects against metabolic stress-induced hepatocyte injury.

To elucidate the potential role of autophagy and the protective effects of Jiang Zhi Granule (JZG) in metabolic stress-induced hepatocyte injury.An and approach was used in this study. HepG2 cells were incubated in culture medium containing palmitate (PA; 0, 0.1, 0.2, 0.3, 0.4 or 0.5 mmol/L) and treated with or without JZG (100 μg/mL) for 24 h or 48 h, and the progression of autophagy was visualized by stable fluorescence-expressing cell lines LC3 and p62. Western blot analyses were performed to examine the expression of LC3-II/LC3-I, p62, mTOR and PI3K, while mitochondrial integrity and oxidative stress were observed by fluorescence staining of JC-1 and reactive oxygen species. C57BL/6 mice were divided into three groups: control group ( = 10), high fat (HF) group ( = 13) and JZG group ( = 13); and, histological staining was carried out to detect inflammation and lipid content in the liver.The cell trauma induced by PA was aggravated in a dose- and time-dependent manner, and hepatic function was improved by JZG. PA had dual effects on autophagy by activating autophagy induction and blocking autophagic flux. The PI3K-AKT-mTOR signaling pathway and the fusion of isolated hepatic autophagosomes and lysosomes were critically involved in this process. JZG activated autophagy progression by either induction of autophagosomes or co-localization of autophagosomes and lysosomes as well as degradation of autolysosomes to protect against PA-induced hepatocyte injury, and protected mitochondrial integrity against oxidative stress in PA-induced mitochondrial dysfunction. In addition, JZG ameliorated lipid droplets and inflammation induced by HF diet , leading to improved metabolic disorder and associated liver injury in a mouse model of non-alcoholic fatty liver disease (NAFLD).Metabolic stress-induced hepatocyte injury exhibited dual effects on autophagy and JZG activated the entire process, resulting in beneficial effects in NAFLD.

Keyword: energy

Characterization and robust nature of newly isolated oleaginous marine yeast Rhodosporidium spp. from coastal water of Northern China.

A total of ten marine yeast strains isolated from Bohai Sea, Northern China were identified to be members of three genera Rhodosporidium, Rhodotorula, and Cryptococcus. Two representative strains Rhodosporidium TJUWZ4 and Cryptococcus TJUWZA11 with high lipid content based on Nile red staining method were further characterized. A wide range of culture conditions (C and N sources, pH, temperature, salinity and C/N ratio) were tested to characterize the biomass and lipid production (yield and productivity) of these strains. Results indicated that Rhodosporidium TJUWZ4 was capable of achieving lipid yield up to 44% and 0.09\xa0g/l-h productivity on glucose and peptone medium at pH 4, 20\xa0°C, 30% salinity, and C/N 80. Three fatty acids, namely oleic (18:1), (C16:0) and linoleic (18:2) were the major intracellular fatty acids, which accounted for 90% of total lipids. With promising features for intracellular lipid accumulation, Rhodosporidium TJUWZ4 is a robust strain with great potentials for application in biodiesel production from renewable feedstocks.

Keyword: energy

Ecological relevance of skeletal fatty concentration and composition in Mediterranean scleractinian corals.

The intra-skeletal fatty concentration and composition of four Mediterranean coral species, namely Cladocora caespitosa, Balanophyllia europaea, Astroides calycularis and Leptopsammia pruvoti, were examined in young and old individuals living in three different locations of the Mediterranean Sea. These species are characterized by diverse levels of organization (solitary or colonial) and trophic strategies (symbiotic or non-symbiotic). Fatty acids have manifold fundamental roles comprehensive of membrane structure fluidity, cell signaling and storage. For all species, except for B. europaea, the intra-skeletal fatty concentration was significantly higher in young individuals than in old ones. Moreover, fatty concentration was higher in colonial corals than in solitary ones and in the symbiotic corals compared to non-symbiotic ones. Analysis by gas chromatography-mass spectrometry (GC-MS) revealed that (16:0) was the most abundant fatty , followed by stearic (18:0) in order of concentration. Oleic (18:1) was detected as the third main component only in skeletons from symbiotic corals. These results suggest that, in the limits of the studied species, intra-skeletal fatty composition and concentration may be used for specific cases as a proxy of level of organization and trophic strategy, and eventually coral age.

Keyword: energy

Hydroxyhydroquinone impairs fat utilization in mice by reducing nitric oxide availability.

Habitual consumption of chlorogenic compounds (CGAs) from coffee increases fat catabolism and reduces body fat; however, the contribution of roasted coffee remains unclear. Hydroxyhydroquinone (HHQ) impairs the vasodilatory and antihypertensive effects of CGAs by reducing nitric oxide (NO) bioavailability. Since HHQ also reduces fat catabolism, we hypothesized that HHQ does so by decreasing NO availability. Therefore, we investigated the effect of HHQ on metabolism in KKAy mice. In HHQ-treated mice, fat oxidation was significantly low and dose-dependent, serum and urinary hydrogen peroxide were high, and plasma NO metabolites and S-nitrosylated liver proteins were low. In HHQ-treated mouse hepatocytes, the palmitate-induced increase in cellular oxygen consumption was negatively affected, and HHQ or L-NAME reduced cellular fatty utilization. In conclusion, HHQ can impair fat utilization by reducing NO availability in mice. Protein S-nitrosylation reduction in liver cells after HHQ consumption may be associated with impaired fatty oxidation.

Keyword: energy

Dyslipidemia impairs mitochondrial trafficking and function in sensory neurons.

Mitochondrial trafficking plays a central role in dorsal root ganglion (DRG) neuronal cell survival and neurotransmission by transporting mitochondria from the neuronal cell body throughout the bundles of DRG axons. In type 2 diabetes (T2DM), dyslipidemia and hyperglycemia damage DRG neurons and induce mitochondrial dysfunction; however, the impact of free fatty acids and glucose on mitochondrial trafficking in DRG neurons remains unknown. To evaluate the impact of free fatty acids compared to hyperglycemia on mitochondrial transport, primary adult mouse DRG neuron cultures were treated with physiologic concentrations of palmitate and glucose and assessed for alterations in mitochondrial trafficking, mitochondrial membrane potential, and mitochondrial bioenergetics. Palmitate treatment significantly reduced the number of motile mitochondria in DRG axons, but physiologic concentrations of glucose did not impair mitochondrial trafficking dynamics. Palmitate-treated DRG neurons also exhibited a reduction in mitochondrial velocity, and impaired mitochondrial trafficking correlated with mitochondrial depolarization in palmitate-treated DRG neurons. Finally, we found differential bioenergetic effects of palmitate and glucose on resting and energetically challenged mitochondria in DRG neurons. Together, these results suggest that palmitate induces DRG neuron mitochondrial depolarization, inhibiting axonal mitochondrial trafficking and altering mitochondrial bioenergetic capacity.-Rumora, A. E., Lentz, S. I., Hinder, L. M., Jackson, S. W., Valesano, A., Levinson, G. E., Feldman, E. L. Dyslipidemia impairs mitochondrial trafficking and function in sensory neurons.© FASEB.

Keyword: energy

Disruption of Lipid Uptake in Astroglia Exacerbates Diet-Induced Obesity.

Neuronal circuits in the brain help to control feeding behavior and systemic metabolism in response to afferent nutrient and hormonal signals. Although astrocytes have historically been assumed to have little relevance for such neuroendocrine control, we investigated whether lipid uptake via lipoprotein lipase (LPL) in astrocytes is required to centrally regulate homeostasis. Ex vivo studies with hypothalamus-derived astrocytes showed that LPL expression is upregulated by oleic , whereas it is decreased in response to or triglycerides. Likewise, astrocytic LPL deletion reduced the accumulation of lipid droplets in those glial cells. Consecutive in vivo studies showed that postnatal ablation of LPL in glial fibrillary acidic protein-expressing astrocytes induced exaggerated body weight gain and glucose intolerance in mice exposed to a high-fat diet. Intriguingly, astrocytic LPL deficiency also triggered increased ceramide content in the hypothalamus, which may contribute to hypothalamic insulin resistance. We conclude that hypothalamic LPL functions in astrocytes to ensure appropriately balanced nutrient sensing, ceramide distribution, body weight regulation, and glucose metabolism.© 2017 by the American Diabetes Association.

Keyword: energy

Enhanced lipid production by Rhodosporidium toruloides using different fed-batch feeding strategies with lignocellulosic hydrolysate as the sole carbon source.

Industrial biotechnology that is able to provide environmentally friendly bio-based products has attracted more attention in replacing petroleum-based industries. Currently, most of the carbon sources used for fermentation-based bioprocesses are obtained from agricultural commodities that are used as foodstuff for human beings. Lignocellulose-derived sugars as the non-food, green, and sustainable alternative carbon sources have great potential to avoid this dilemma for producing the renewable, bio-based hydrocarbon fuel precursors, such as microbial lipid. Efficient bioconversion of lignocellulose-based sugars into lipids is one of the critical parameters for industrial application. Therefore, the\xa0fed-batch cultivation, which is a common method used in industrial applications, was investigated to achieve a high cell\xa0density culture along with high lipid yield and productivity.In this study, several fed-batch strategies were explored to improve lipid production using lignocellulosic hydrolysates derived from corn stover. Compared to the batch culture giving a lipid yield of 0.19\xa0g/g, the dissolved-oxygen-stat feeding mode increased the lipid yield to 0.23\xa0g/g and the lipid productivity to 0.33\xa0g/L/h. The pulse feeding mode further improved lipid productivity to 0.35\xa0g/L/h and the yield to 0.24\xa0g/g. However, the highest lipid yield (0.29\xa0g/g) and productivity (0.4\xa0g/L/h) were achieved using an automated online sugar control feeding mode, which gave a dry cell weight of 54\xa0g/L and lipid content of 59\xa0% (w/w). The major fatty acids of the lipid derived from lignocellulosic hydrolysates were predominately and oleic , which are similar to those of conventional oilseed plants.Our results suggest that the fed-batch feeding strategy can strongly influence the lipid production. The online sugar control feeding mode was the most appealing strategy for high cell density, lipid yield, and lipid productivity using lignocellulosic hydrolysates as the sole carbon source.

Keyword: energy

Changes of albumin secondary structure after binding. FT-IR spectroscopic study.

Albumin is an universal transport protein. Plasma pool of free fatty acids arising from triglyceride hydrolysis, critical in metabolism and etiology of metabolic disorders is transported by albumin. According to various studies albumin has from seven to nine binding sites with diverse affinity to long chain fatty acids. X-ray diffraction crystallography measurements have provided data only for pure human serum albumin or albumin with fully saturated binding sites. These results have shown that amount of \uf061-helices is higher after fatty acids binding. Molecular mechanics simulations suggest that binding of fatty acids in two high-affinity sites leads to major conformational changes in albumin structure. The aim of this research was to investigate albumin secondary structure upon gradually increasing fatty acids to protein mole ratio.Fourier transform infrared spectroscopy was applied to study changes of bovine serum albumin (as an analogue of human serum albumin) \uf061-helical structures after binding in a range of 0-20 : albumin molar ratios representing pure protein, partial, full saturation and excess binding sites capacity.Amount of \uf061-helices was increasing along with the amount of : bovine serum albumin molar ratio and reached maximum value around 2 mol/mol.Our studies confirmed molecular mechanics simulations and crystallographic studies. binding in two high-affinity sites leads to major structural changes, filling another sites only slightly influenced bovine serum albumin secondary structure. The systematic study of fatty acids and albumin interactions, using an experimental model mimicking metabolic disorders, may results in new tools for personalized nanopharmacotherapy.

Keyword: energy

Exposure of bovine oocytes and embryos to elevated non-esterified fatty concentrations: integration of epigenetic and transcriptomic signatures in resultant blastocysts.

Metabolic stress associated with negative balance in high producing dairy cattle and obesity in women is a risk factor for decreased fertility. Non-esterified fatty acids (NEFA) are involved in this pathogenesis as they jeopardize oocyte and embryo development. Growing evidence indicates that maternal metabolic disorders can disturb epigenetic programming, such as DNA methylation, in the offspring. Oocyte maturation and early embryo development coincide with methylation changes and both are sensitive to adverse environments. Therefore, we investigated whether elevated NEFA concentrations affect establishment and maintenance of DNA methylation in oocytes and embryos, subsequently altering transcriptomic profiles and developmental competence of resultant blastocysts.Bovine oocytes and embryos were exposed to different NEFA concentrations in separate experiments. In the first experiment, oocytes were matured in vitro for 24\xa0h in medium containing: 1) physiological ("BASAL") concentrations of oleic (OA), (PA) and stearic (SA) or 2) pathophysiological ("HIGH COMBI") concentrations of OA, PA and SA. In the second experiment, zygotes were cultivated in vitro for 6.5\xa0days under BASAL or HIGH COMBI conditions. Developmental competence was evaluated by assessing cleavage and blastocyst rate. Overall gene expression and DNA methylation of resultant blastocysts were analyzed using microarray. DNA methylation data were re-evaluated by pyrosequencing. HIGH COMBI-exposed oocytes and embryos displayed a lower competence to develop into blastocysts compared to BASAL-exposed counterparts (19.3% compared to 23.2% and 18.2% compared to 25.3%, respectively) (P\u2009<\u20090.05). HIGH COMBI-exposed oocytes and embryos resulted in blastocysts with altered DNA methylation and transcriptomic fingerprints, compared to BASAL-exposed counterparts. Differences in gene expression and methylation were more pronounced after exposure during culture compared to maturation suggesting that zygotes are more susceptible to adverse environments. Main gene networks affected were related to lipid and carbohydrate metabolism, cell death, immune response and metabolic disorders.Overall, high variation in methylation between blastocysts made it difficult to draw conclusions concerning methylation of individual genes, although a clear overview of affected pathways was obtained. This may offer clues regarding the high rate of embryonic loss and metabolic diseases during later life observed in offspring from mothers displaying lipolytic disorders.

Keyword: energy

Elevated free fatty acids affect bovine granulosa cell function: a molecular cue for compromised reproduction during negative balance.

High-yielding dairy cows postpartum face the challenge of negative balance leading to elevated free fatty acids levels in the serum and follicular fluid thus affecting the ovarian function. Here, we investigated effects of physiological concentrations of (PA), stearic (SA) and oleic (OA) on the viability, steroid production and gene expression in a bovine granulosa cell (GC) culture model. Treatment with individual and combined fatty acids increased the CD36 gene expression, while no significant apoptotic effects were observed. Both PA and SA significantly upregulated the expression of FSHR, LHCGR, CYP19A1, HSD3B1, CCND2 and increased 17β-estradiol (E2) production, while OA downregulated the expression of these genes and reduced E2. Interestingly, STAR was equally downregulated by all fatty acids and combination treatment. E2 was significantly reduced after combination treatment. To validate the effects of OA, in vivo growing dominant follicles (10-19 mm) were injected with bovine serum albumin (BSA) with/without conjugated OA. The follicular fluid was recovered 48 h post injection. As in our in vitro model, OA significantly reduced intrafollicular E2 concentrations. In addition, expression of CD36 was significantly up- and that of CYP19A1 and STAR significantly downregulated in antral GC recovered from aspirated follicles. The ovulation rates of OA-injected follicles tended to be reduced. Our results indicate that elevated free fatty concentrations specifically target functional key genes in GC both in vitro and in vivo. Suggestively, this could be a possible mechanism through which elevated free fatty acids affect folliculogenesis in dairy cows postpartum.

Keyword: energy

Detection of organic compounds in impact glasses formed by the collision of an extraterrestrial material with the Libyan Desert (Africa) and Tasmania (Australia).

Impact glasses are rich silica melted formed at high temperature and pressure by the impact of an extraterrestrial body on Earth. Here, Libyan Desert glasses (LDGs) and Darwin glasses (DGs) were studied. Two non-destructive analytical techniques were used to detect and characterize organic compounds present in their inclusions: Raman spectroscopy and scanning electron microscopy coupled to -dispersive X-ray spectroscopy (SEM-EDS). Phytoliths, humboldtine, , myristic , oleic , 4-methyl phthalic , and S-H stretching vibrations of amino acids were identified. The presence of these particular organic compounds in such materials has not been reported so far, providing information about (a) the ancient matter of the area where the impact glasses were formed, (b) organic matter belonging to the extraterrestrial body which impacted on the Earth, or (c) even to current plant or bacterial life, which could indicate an active interaction of the LDG and DG with the surrounding environment. Moreover, the identification of fullerene allowed us to know a pressure (15\xa0GPa) and temperatures (670\xa0K or 1800-1900\xa0K) at which samples could be subjected.

Keyword: energy

Novel Mechanisms Modulating Palmitate-Induced Inflammatory Factors in Hypertrophied 3T3-L1 Adipocytes by AMPK.

A growing body of evidence indicates that AMP-activated protein kinase (AMPK) contributes to not only metabolic homeostasis but also the inhibition of inflammatory responses. However, the underlying mechanisms remain unclear. To elucidate the role of AMPK, in this study, we observed the effects of AMPK activation on monocyte chemoattractant protein-1 (MCP-1) release in mature 3T3-L1 adipocytes.We observed signal transduction pathways regulating MCP-1, which increased in obese adipocytes, in an model of hypertrophied 3T3-L1 adipocytes preloaded with palmitate.Palmitate-preloaded cells exhibited significant increase in MCP-1 release and triglyceride (TG) deposition. Increased MCP-1 release and TG deposition were significantly decreased by an AMPK activator. In addition, the AMPK activator not only markedly diminished MCP-1 secretion but also augmented phosphorylation of nuclear factor-B (NF-B) and extracellular signal-regulated kinase (ERK) 1/2. In contrast, MCP-1 release suppression was abolished by the AMPK inhibitor compound C and the MEK inhibitor U0126.MCP-1 release from hypertrophied adipocytes is suppressed by AMPK activation through the NF-B and ERK pathways. These findings provide evidence that AMPK plays a crucial role in ameliorating obesity-induced inflammation.

Keyword: energy

α-Ionone attenuates high- diet-induced skeletal muscle wasting in mice via activation of cAMP signaling.

Skeletal muscle atrophy is a common and debilitating condition that lacks a pharmacological therapy. Our aim was to investigate the potential of α-ionone, a naturally occurring flavoring agent, in preventing muscle atrophy and to delineate the mechanisms involved. We found that α-ionone not only stimulated myogenesis but also attenuated -induced atrophy of cultured skeletal myotubes, as evidenced by an increased myotube diameter and length, fusion index, and cellular protein content. These beneficial actions of α-ionone were abrogated by cAMP inhibitor. The antiatrophic effects of α-ionone on cultured myotubes were confirmed in a corresponding mouse model. The skeletal muscle mass, muscle protein content, myofiber diameter, and muscle strength were greater in α-ionone-treated mice than in untreated animals fed high- diet. Furthermore, α-ionone increased cAMP concentration and enhanced its downstream PKA-CREB signaling in skeletal muscle of mice fed high- diet. Thus, α-ionone is a promising agent that may enhance skeletal muscle mass and strength.

Keyword: fat metabolism

Dietary zerumbone from shampoo ginger: new insights into its antioxidant and anticancer activity.

The dietary sesquiterpene dienone zerumbone (ZER) selectively targets cancer cells, inducing mitochondrial dysfunction and apoptosis, and protects non-cancerous cells towards oxidative stress and insult. This study examines the in vitro effects of ZER on peroxidation in biological systems (cholesterol and phospholipid membrane oxidation) and explores its antitumor action in terms of its ability to modulate cancer cell profile. Evaluation of the antioxidant activity of ZER showed that this compound is unable to trap lipoperoxyl radicals per se. ZER significantly modulated the total and fatty profiles in cancer cells, inducing marked changes in the phospholipid/cholesterol ratio, with significant decreases in the levels of oleic and acids and a marked increase of stearic . Cell-based fluorescent measurements of intracellular membranes and droplets using the Nile Red staining technique showed that in cancer cells, ZER induced significant accumulation of cytosolic droplets and altered cell membrane organization/protein dynamics, depolarizing the mitochondrial membranes and inducing apoptosis and alteration of nuclear morphology. The modulatory activity of ZER on the total and fatty profiles and droplets may therefore represent another possible mechanism of its anticancer properties.

Keyword: fat metabolism

Overexpression of Acyl-ACP Thioesterases, and , Induce Distinct Gene Expression Reprogramming in Developing Seeds of .

We examined the substrate preference of acyl-ACP thioesterases, and , and gene expression changes associated with the modification of composition in the seed, using transgenic plants overexpressing or under the control of a seed-specific promoter. seeds contained a higher level of total saturated fatty (FA) content, with 4.3 times increase in 16:0 , whereas seeds showed approximately 3% accumulation of 10:0 and 12:0 medium-chain FAs, and a small increase in other saturated FAs, resulting in higher levels of total saturated FAs. RNA-Seq analysis using entire developing pods at 8, 25, and 45 days after flowering (DAF) showed up-regulation of genes for β-ketoacyl-acyl carrier protein synthase I/II, stearoyl-ACP desaturase, oleate desaturase, and linoleate desaturase, which could increase unsaturated FAs and possibly compensate for the increase in 16:0 at 45 DAF in transgenic plants. In transgenic plants, many putative chloroplast- or mitochondria-encoded genes were identified as differentially expressed. Our results report comprehensive gene expression changes induced by alterations of seed FA composition and reveal potential targets for further genetic modifications.

Keyword: fat metabolism

Irisin Ameliorates Glucolipotoxicity-Associated β-Cell Dysfunction and Apoptosis via AMPK Signaling and Anti-Inflammatory Actions.

Islet metabolic disorder and inflammation contribute to the pathogenesis and progression of type 2 diabetes mellitus (T2DM). Irisin is a recently identified adipomyokine with protective effects on metabolic homeostasis and inflammation-suppressing effects in hepatic and vascular cells. The present study examined the effects of irisin on and inflammation in β cells under glucolipotoxic conditions.Rat INS-1E β cells and islets isolated from C57BL/6 mice were incubated in glucolipotoxic conditions with or without irisin. Intracellular contents and lipogenic gene expression were determined by enzymatic colorimetric assays and real-time PCR, respectively. Inflammatory status was evidenced by Western blot analysis for the phosphorylation of nuclear factor-κB (NF-κB) p65 and real-time PCR analysis for the expression of pro-inflammatory genes.Irisin reversed glucolipotoxicity-induced intracellular non-esterified fatty (NEFA) and triglyceride accumulation, suppressed associated elevations in lipogenic gene expression, and phosphorylated acetyl-CoA-carboxylase (ACC) in INS-1E cells. These demonstrated effects were dependent on irisin-activated adenosine monophosphate-activated protein kinase (AMPK). Meanwhile, AMPK signaling mediated the protective effects of irisin on INS-1E cell insulin secretory ability and survival as well. Additionally, irisin inhibited phosphorylation of NF-κB p65 while decreasing the expression of pro-inflammatory genes in INS-1E cells under glucolipotoxic conditions. Moreover, irisin also improved insulin secretion, inhibited apoptosis, and restored β-cell function-related gene expression in isolated mouse islets under glucolipotoxic conditions.Irisin attenuated excessive lipogenesis in INS-1E cells under glucolipotoxic state through activation of AMPK. Irisin also suppressed overnutrition-induced inflammation in INS-1E cells. Our findings implicate irisin as a promising therapeutic target for the treatment of islet metabolic disorder and islet inflammation in T2DM.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fat metabolism

Compound C Protects Mice from HFD-Induced Obesity and Nonalcoholic Fatty Liver Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high- diet- (HFD-) induced obesity and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce obesity and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. Liver histology was observed. The expression levels of genes related to and proinflammation in liver tissue were examined. NLRP3 inflammasome expression in liver tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in body-weight gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and insulin tolerance test (ITT) showed that compound C alleviated insulin resistance. Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased fatty synthesis genes, while increased fatty oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related metabolic disorders.

Keyword: fat metabolism

[The becoming of reactions of lipolysis in phylogenesis. The and oleic triglycerides as substrates. Insulin, condition of normolipemia and formation of hyper lipoproteinemia type IIb, IV and V].

According to phylogenetic theory of general pathology, when living in ocean all were carnivorous (piscivorous) fatty acids transferring to cells in form of non-polar triglycerides nitially began apoB-48 chylomicrons, continued lipoproteins of very low and low density and fnalized its apoB-100 endocytosis. The fatty acids are transferred by chylomicrons + lipoproteins of very low density + lipoproteins of low density and non-polar triglycerides are hydrolyzed by hepatic glycerolhydrogenase and co-enzyme apoC-III; according WHO classifcation, hyperlipoproteinemia corresponds to type V. On land, in herbivorous who are not yet synthesized insulin, apoB-48 and chylomicrons left process of non-polar triglycerides transferring. In lipoproteins of very low density and lipoproteins of low density, the carnivorous transfer exogenous non-polar triglycerides. The herbivorous also transfer non-polar triglycerides though synthesized by hepatocytes from glucose endogenically. In herbivorous, transferring of non-polar triglycerides prior to synthesis of insulin is forming apoB-100 in composition of lipoproteins of very low density and lipoproteins of low density. The hydrolysis of non-polar triglycerides in lipoproteins of very low density is activated by hepatic glycerol hydrogenase and apoC-III; cells absorb lipoproteins of low density by means of apoB-100 endocytosis. The content on lipoproteins in blood plasma under electrophoresis of lipoproteins corresponds to hepatic glycerol hydrogenase type IIb. In frst and second types of fatty acids transferring in form of triglycerides to lipoproteins of very low density + lipoproteins of low density predominate fatty , triglycerides of the same name and of fatty acids in vivo. The insulin initiated the third type of transferring of oleic fatty by now to insulin-depended cells only in oleic lipoproteins of very low density; hydrolysis of oleic triglycerides is activated by late in phylogenesis post-heparin hepatic glycerol hydrogenase and apoC-II cofactor. The dynamic apoE is actively bound by apoB-100 forming apoE/B-100 ligand. At later stages of phylogenesis insulin formed fatty acids transferring in form of oleic triglycerides in lipoproteins of very low density of the same name without forming of oleic lipoproteins of low density; the electrophoregram of lipoproteins reflects absence of hepatic glycerol hydrogenase. In phylogenesis three types of fatty acids transferring to triglycerides in composition of lipoproteins formed sequentially: 1) chylomicrons + lipoproteins of very low and density + lipoproteins of low density; 2) lipoproteins of very low density + lipoproteins of low density; 3) only in lipoproteins of very low density. The frst one is specifc to piscivorous (carnivorous) while living in ocean. The second one is implemented by herbivorous while they didn\'t begin to synthesize insulin and hepatocytes not yet transform all endogenous fatty into oleic fatty . Insulin initiated: a) transferring of oleic fatty acids to lipoproteins of very low density without forming oleic lipoproteins of low density; b) highly effective oleic of fatty acids in vivo: c) becoming of biological function of locomotion. The aphysiological induction by substrate, surplus of fatty acids in food initiate negative alterations in composition of lipoproteins in opposite direction than in case of phylogenesis. When homo sapiens, herbivorous in phylogenesis, begins to misuse carnivorous (meat) food then instead of normolipoproteinemia in blood plasma under electrophoresis of lipoproteins one can initially detect transitory hyperlipoproteinemia type IV and then prolonged hyperlipoproteinemia type IIb. If patient factually passes on to carnivorous diet then hyperlipoproteinemia type V is developing. If content of exogenous fatty in food surpasses physiological capacities of its transferring in oleic triglycerides as palmitoyl-oleyl-palmitate glycerol, triglycerides as oleyl-palmitoyl-palmitate glycerol begin to form and epigenetically aphysiological non-ligand lipoproteins of very low density → lipoproteins of low density are formed. Their circulation in blood is a cause of hypertriglyceridemia, higher level of cholesterol-lipoproteins of low density, compensatory increasing of apoC-III. Then occurs induced by substrate formation of hyperlipoproteinemia initially of type IV, then of type IIb and fnally of type V. The pathogenesis of atherosclerosis and atheromotosis is activated when homo sapiens, herbivorous in phylogenesis, begin to misuse carnivorous food affecting biological functions of trophology, reaction of exotrophy (external nutrition), function of homeostasis, endoecology and function of adaptation. The formation of if fatty acids instead of oleic one is a cause of chronic defciency of energy and ATP synthesis in vivo. Insulin activates absorption of glucose by cells with purpose to use it for synthesis of oleic fatty acids. In the frst place, insulin regulates in vivo of fatty acids and only in second place of glucose.

Keyword: fat metabolism

Flavone glycosides from Sicyos angulatus and their inhibitory effects on hepatic accumulation.

A library of extracted natural materials (Korea Bioactive Natural Material Bank) have been screened to discover candidates for the treatment of non-alcoholic liver disease (NAFLD), and the 70% ethanol extract of Sicyos angulatus was found to inhibit hepatic accumulation. Bioassay-guided fractionation of this bioactive extract yielded five previously undescribed flavonoid glycosides and one previously undescribed flavonolignan glycoside along with seven known flavonoid glycosides. The chemical structures of these compounds were elucidated by a combination of extensive spectroscopic analysis, including MS, NMR and UV techniques. Eight compounds of all isolated compounds showed inhibitory effects on the accumulation induced by high concentrations of and glucose in HepG2 cells. Four selected compounds were tested for content in a dose-dependent manner (10, 20 and 40\u202fμM), and among those compounds, kaempferol 3-O-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside showed the strongest inhibition of hepatic production in HepG2 cells. In an oil-red O staining assay, five compounds were shown to reduce hepatic accumulation better than what was observed in the vehicle control group. The present study suggests a new class of chemical entities for developing bioactive agents for the treatment of diseases caused by accumulation in the liver.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Digestibility of amino in full- canola seeds, canola meal, and canola expellers fed to broiler chickens and pigs.

Canola products including full- canola seeds (FFCS), canola meal (CM), and canola expellers (CE) have been used in diets for both broiler chickens and pigs. However, their ability to utilize the AA in canola products might be different from each other. Therefore, this study was conducted to compare the apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of CP and AA in broiler chickens and growing pigs fed FFCS, CM, and CE. Three diets were prepared to contain FFCS, CM, or CE as a sole source of nitrogen. A nitrogen-free diet was prepared. In Exp. 1, a total of 272 twenty-one-day-old male broiler chickens with an initial BW of 932 ± 80.6 g were assigned to diets in a randomized complete block design with BW as a blocking factor. After 5 d of feeding, birds were euthanized by sodium pentobarbital, and ileal digesta samples were collected from distal two-third of the ileum. In Exp. 2, 16 barrows were surgically fitted with T-cannulas at the distal ileum. After 8-d recovery period, pigs (initial BW = 18.9 ± 1.17 kg) were divided into 4 blocks based on BW and assigned to a quadruplicate 4 × 2 incomplete Latin Square design with 4 diets and 2 periods. Each period consisted of 5-d adaptation and 2-d ileal digesta collection periods. Data were analyzed as a 2 × 3 factorial treatment arrangement with effects of species (broiler chickens or pigs) and diets (FFCS, CM, or CE). There were interactions (P < 0.05) between species and experimental diets in the AID of all indispensable AA except for Lys. The AID of indispensable AA in FFCS for broiler chickens was greater (P < 0.05) than for pigs. Broiler chickens also had greater (P < 0.05) AID of Arg, His, Leu, Phe, and Val in CM compared with pigs; however, there were no differences in the AID of indispensable AA in CE between broiler chickens and pigs. The basal ileal endogenous losses of CP and AA, except Trp, in pigs were greater (P < 0.05) than in broiler chickens. There were also interactions (P < 0.05) between species and experimental diets in the SID of all indispensable AA except for Lys. Broiler chickens fed the diet containing FFCS had greater (P < 0.05) SID of indispensable AA compared with pigs fed the same diet; however, the SID of indispensable AA in CM or CE were not different between broiler chickens and pigs. In conclusion, differences in digestibility of AA in canola products were affected by nonruminant animal species.

Keyword: fat metabolism

Palmitoylethanolamide Promotes a Proresolving Macrophage Phenotype and Attenuates Atherosclerotic Plaque Formation.

Objective- Palmitoylethanolamide is an endogenous fatty mediator that is synthetized from membrane phospholipids by N-acyl phosphatidylethanolamine phospholipase D. Its biological actions are primarily mediated by PPAR-α (peroxisome proliferator-activated receptors α) and the orphan receptor GPR55. Palmitoylethanolamide exerts potent anti-inflammatory actions but its physiological role and promise as a therapeutic agent in chronic arterial inflammation, such as atherosclerosis remain unexplored. Approach and Results- First, the polarization of mouse primary macrophages towards a proinflammatory phenotype was found to reduce N-acyl phosphatidylethanolamine phospholipase D expression and palmitoylethanolamide bioavailability. N-acyl phosphatidylethanolamine phospholipase D expression was progressively downregulated in the aorta of apolipoprotein E deficient (ApoE) mice during atherogenesis. N-acyl phosphatidylethanolamine phospholipase D mRNA levels were also downregulated in unstable human plaques and they positively associated with smooth muscle cell markers and negatively with macrophage markers. Second, ApoE mice were fed a high- diet for 4 or 16 weeks and treated with either vehicle or palmitoylethanolamide (3 mg/kg per day, 4 weeks) to study the effects of palmitoylethanolamide on early established and pre-established atherosclerosis. Palmitoylethanolamide treatment reduced plaque size in early atherosclerosis, whereas in pre-established atherosclerosis, palmitoylethanolamide promoted signs of plaque stability as evidenced by reduced macrophage accumulation and necrotic core size, increased collagen deposition and downregulation of M1-type macrophage markers. Mechanistically, we found that palmitoylethanolamide, by activating GPR55, increases the expression of the phagocytosis receptor MerTK (proto-oncogene tyrosine-protein kinase MER) and enhances macrophage efferocytosis, indicative of proresolving properties. Conclusions- The present study demonstrates that palmitoylethanolamide protects against atherosclerosis by promoting an anti-inflammatory and proresolving phenotype of lesional macrophages, representing a new therapeutic approach to resolve arterial inflammation.

Keyword: fat metabolism

Dietary sn-2 triacylglycerols reduced faecal lipids, calcium contents and altered in Sprague-Dawley rats.

In this study, the impact of dietary sn-2 triacylglycerol (sn-2 PTAG) on faecal lipids, calcium excretion and metabolic alternation was investigated in Sprague-Dawley (SD) rats fed with high- diet containing either palm olein (PO, sn-2 (PA) of 14.8%), sn-2 PTAG50 (sn-2\u2009PA of 56.4%) or sn-2 PTAG70 (sn-2\u2009PA of 72.4%), respectively. After 4-week feeding period, SD rats fed with sn-2 PTAGs showed reduced faecal soap fatty acids, neutral and calcium excretion compared to those of PO-fed rats, whereas a significant difference was only observed for the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Moreover, dietary sn-2 PTAG70 also showed a significant effect on decreasing serum triacylglycerol (TAG) level, reducing perirenal adipocyte size and regulating in small intestine and perirenal adipose tissue of SD rats. Significantly increased mRNA levels of genes involved in intestinal anabolism as well as catabolism were both observed in the sn-2 PTAG70-fed rats (p\u2009<\u2009.05). Meanwhile, dietary sn-2 PTAG70 also significantly up-regulated lipolysis, mitochondrial fatty oxidation and thermogenesis-related gene and protein levels in perirenal adipose tissue, which might be correlated with the reduced perirenal adipocyte size. Taken together, our findings indicated that sn-2 PTAG70 may have some beneficial effects on intestinal utilisation and metabolic activity for energy supply in visceral adipose tissue.

Keyword: fat metabolism

Probing the Global Cellular Responses to Lipotoxicity Caused by Saturated Fatty Acids.

Excessive levels of saturated fatty acids are toxic to cells, although the basis for this lipotoxicity remains incompletely understood. Here, we analyzed the transcriptome, lipidome, and genetic interactions of human leukemia cells exposed to palmitate. Palmitate treatment increased saturated glycerolipids, accompanied by a transcriptional stress response, including upregulation of the endoplasmic reticulum (ER) stress response. A comprehensive genome-wide short hairpin RNA (shRNA) screen identified >350 genes modulating lipotoxicity. Among previously unknown genetic modifiers of lipotoxicity, depletion of RNF213, a putative ubiquitin ligase mutated in Moyamoya vascular disease, protected cells from lipotoxicity. On a broader level, integration of our comprehensive datasets revealed that changes in di-saturated glycerolipids, but not other classes, are central to lipotoxicity in this model. Consistent with this, inhibition of ER-localized glycerol-3-phosphate acyltransferase activity protected from all aspects of lipotoxicity. Identification of genes modulating the response to saturated fatty acids may reveal novel therapeutic strategies for treating metabolic diseases linked to lipotoxicity.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

The effect of oak tannin (Quercus robur) and hops (Humulus lupulus) on dietary nitrogen efficiency, methane emission, and milk fatty composition of dairy cows fed a low-protein diet including linseed.

The objective of this study was to test the effects of inclusion of hop pellets (HP) and oak tannin extracts (OT) alone or in combination on N efficiency, methane (CH) emission, and milk production and composition in 2 experiments with dairy cows fed low-N rations supplemented with linseed. In both experiments, 6 lactating Holstein cows were assigned to 3 dietary treatments in a 3 × 3 duplicated Latin square design (21-d periods). Cows were fed a total mixed ration at a restricted level to meet their nutrient requirements. In experiment 1, 169 g dry matter (DM) of OT or 56 g DM of HP was included separately in the control diet (C1). In experiment 2, the additives were included together (OT-HP) in the control diet (C2) similar to C1. Diet C2 was compared with a control without linseed (C0). In experiment 1, the supplementation of the control diet with OT decreased urinary N excretion by 12%. In experiment 2, the combination of OT and HP decreased urinary N by 7%. Oak tannin extracts and HP alone or in combination did not influence the daily enteric CH production of cows. Cows fed diet C0 produced 17% more enteric CH daily than those fed diet C2. Intake of diet C2, which contained 6.7% extruded linseed on a DM basis (experiment 2), decreased the sum of 6:0 to 14:0 fatty acids (-16%) and (-26%) and increased the stearic (+50%), oleic (+36%), vaccenic (trans-11 18:1; +285%), rumenic (cis-9,trans-11 18:2; +235%), and α-linolenic (+100%) in milk . The supplementation of diet C2 with the OT-HP mixture further improved the milk\'s fatty composition. Intake of the OT alone increased α-linolenic by 17.7% (experiment 1). The results of this study show that at the economically acceptable dose we tested, hops had no effect on urinary N excretion, CH emission, milk production, and milk composition. By contrast, supplementation of diets with oak tannin extract can be considered for reducing urinary N excretion. The combination of oak tannin and hops had no more effect than oak tannin alone except on the milk fatty profile, which was favorably influenced from a nutritional point of view.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Metabolic dependence of cyclosporine\xa0A on cell proliferation of human non‑small cell lung cancer A549 cells and its implication in post‑transplant malignancy.

Cyclosporine\xa0A (CsA), a widely used immunosuppressant to prevent organ transplant rejection, is associated with an increased cancer risk following transplantation, particularly in the lung. However, the underlying mechanisms remain unclear. In the present study, using human non‑small cell lung cancer A549 cells, it was determined that CsA (0.1 or 1\xa0µM) promoted cell proliferation with glucose alone as the energy source. CsA treatment increased the phosphorylation of protein kinase\xa0B (Akt) and consequently the expression of Cyclin\xa0D1. Inhibiting Akt signaling with the phosphatidylinositol 3‑kinase inhibitor wortmannin prevented this effect. Mechanistically, CsA treatment increased reactive oxygen species (ROS) generation, and the intracellular ROS scavenger N‑acetyl‑cysteine (NAC) attenuated CsA‑induced cell proliferation as well as the activation of Akt/Cyclin\xa0D1 signaling. However, notably, it was demonstrated that CsA treatment decreased cell proliferation and Akt phosphorylation under normal loading. Further investigation indicated that induced excessive generation of ROS, while CsA treatment further stimulated this ROS production. Scavenging intracellular ROS with NAC attenuated the CsA‑mediated inhibition of cell proliferation. Collectively, the results indicated a pleiotropic effect of CsA in the regulation of A549\xa0cell proliferation under different metabolic conditions. This indicated that CsA administration may contribute to increased post‑transplant cancer risk in organ recipients.

Keyword: fat metabolism

Yeast Smell Like What They Eat: Analysis of Volatile Organic Compounds of in Growth Media Supplemented with Different Lipids.

is part of the human skin microbiota. Its volatile organic compounds (VOCs) possibly contribute to the characteristic odour in humans, as well as to microbiota interaction. The aim of this study was to investigate how the composition of the liquid medium influences the production of VOCs. Growth was performed in four media: (1) mDixon, (2) oleic (OA), (3) oleic + (OA+PA), and (4) (PA). The profiles of the VOCs were characterized by HS-SPME/GC-MS in the exponential and stationary phases. A total number of 61 VOCs was found in , among which alkanes, alcohols, ketones, and furanic compounds were the most abundant. Some compounds previously reported for (γ-dodecalactone, 3-methylbutan-1-ol, and hexan-1-ol) were also found. Through our experiments, using univariate and multivariate unsupervised (Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA)) and supervised (Projection to Latent Structures Discriminant Analysis (PLS-DA)) statistical techniques, we have proven that each tested growth medium stimulates the production of a different volatiles profile in . Carbon dioxide, hexan-1-ol, pentyl acetate, isomer5 of methyldecane, dimethyl sulphide, undec-5-ene, isomer2 of methylundecane, isomer1 of methyldecane, and 2-methyltetrahydrofuran were established as differentiating compounds among treatments by all the techniques. The significance of our findings deserves future research to investigate if certain volatile profiles could be related to the beneficial or pathogenic role of this yeast.

Keyword: fat metabolism

Changes of the Fatty Profile in Erythrocyte Membranes of Patients following 6-Month Dietary Intervention Aimed at the Regression of Nonalcoholic Fatty Liver Disease (NAFLD).

Nonalcoholic fatty liver disease (NAFLD) is closely related to the disorders of fatty acids. The pathogenesis of the disease includes an increased concentration of FFA in blood, an increase in the biosynthesis of fatty acids, and disorders in the process of -oxidation.The aim of the study was to analyze the fatty acids in erythrocyte membranes among 55 patients with NAFLD who were subjected to a 6-month dietary intervention in order to reduce fatty liver.Basic anthropometric and biochemical measurements were performed. The profile of fatty acids was measured in the membranes of erythrocytes and analyzed by gas chromatography. The dietary compliance was evaluated using 72-diary questionnaires, anthropometric measurements.With the reduction of fatty liver (p<0.01), the patients\' biochemical and anthropometric parameters were significantly improved. A significant decrease in the concentration of alanine aminotransferase (p<0.01) and asparagine aminotransferase (p<0.01) was observed, along with a decrease in the amount of insulin (p<0.05) and insulin resistance (p<0.05). Significant changes in terms of the fatty profile were observed among patients who followed the dietary intervention. There was a noticeable tendency in terms of the reduction (p<0.055) and a significant reduction of stearic (p<0.05). Significant changes in the profile of fatty acids were also associated with the reductionof palmitoleic (p<0.05) and oleic acids (p<0.05). Another statistically significant change observed was the increase in polyunsaturated fatty acids. In particular (p<0.01) the rise of eicosapentaenoic (p<0.055) and docosahexaenoic acids (p<0.55) was noted.The profile of fatty acids turned out to be a potential biomarker of the liver changes during NAFLD regression. Further research is needed to fully elucidate the usefulness and applicability of our findings in the management of NAFLD.

Keyword: fat metabolism

11β-hydroxysteroid dehydrogenase type 1 inhibitor attenuates high- diet induced cardiomyopathy.

High- diet (HFD) induces cardiac hypertrophy; however, the underlying cellular and molecular mechanisms are yet unclear. In the present study, we investigated the roles of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), an amplifier of local glucocorticoid activity, in the pathogenesis of cardiac dysfunction.Male Wistar rats were fed normal chow diet (NC) or HFD and examined the cardiac remolding and functional alteration by echocardiography and histology. Primary neonatal rat ventricular cardiomyocytes (NRCMs) treated with (PA) or infected by lentivirus were used for identifying the role by 11β-HSD1 in cardiac hypertrophy. Genome microarray of NRCMs was performed to further reveal the mechanism underlying cardiac dysfunction. induced hypertrophy in NRCMs that upregulated 11β-HSD1 expression in cardiomyocytes, which led to a significant enlargement in the cell size and expression of cardiac hypertrophy-specific genes. Conversely, a remarkable decrease in cardiomyocytes size was detected in either BVT.2733 (a selective inhibitor of 11β-HSD1)-treated or 11β-HSD1-deficient NRCMs. Furthermore, both glucocorticoid receptor (GR) antagonist RU486 and mineralocorticoid receptor (MR) antagonist spironolactone markedly attenuated the 11β-HSD1-induced cardiomyocytes hypertrophy. Genome microarray revealed that cAMP and calcium signaling pathways are potential downstream signaling pathways regulated by 11β-HSD1 in cardiomyocytes hypertrophy. Similar to in vitro results, BVT.2733 strikingly attenuated cardiac hypertrophy and improved cardiac function in HFD-fed rats.11β-HSD1 acts as an important regulator that controls the cardiac remolding via both GR and MR and the pharmacological inhibition of 11β-HSD1 could be a new therapeutic approach in preventing HFD-induced cardiac hypertrophy.Copyright © 2018. Published by Elsevier Ltd.

Keyword: fat metabolism

Comprehensive characterization of nanostructured carriers using laboratory and synchrotron X-ray scattering and diffraction.

The development of nanoparticles requires knowledge on the crystalline structure, polymorphic transitions and -drug interactions. This study aimed at introducing advanced techniques to characterize nanostructured carriers (NLC) comprising , oleic , stabilizer and Domperidone. Crystallinity of single components and mixtures was investigated by laboratory Small Angle X-ray Scattering (SAXS). NLC were studied with laboratory Small and Wide Angle X-ray Scattering (SWAXS). Photon Correlation Spectroscopy and Freeze Fracture Transmission Electron Microscopy were used to monitor particle size, zeta potential and shape. Stability of NLC was investigated using synchrotron X-ray Diffraction (XRD) and SAXS and laboratory SAXS. showed a lamellar structure (polymorph C), which was still present after particle preparation. Spherical 300\u202fnm-sized particles with zeta potential values above -30\u202fmV were obtained and Domperidone was incorporated in its amorphous form. During storage, no differences in synchrotron XRD spectra were seen. However, laboratory SAXS measurements showed a second lamellar structure, identified as polymorph B. Synchrotron SAXS temperature scans confirmed that polymorph B did not affect the morphology of the encapsulated drug or the shape of NLC. These results highlight the unique capabilities of laboratory and synchrotron X-ray Scattering and Diffraction for improved structural characterization of nanoparticles.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in energy . However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high- diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress lipogenesis. In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic fatty liver disease.

Keyword: fat metabolism

Relationship between selenium removal efficiency and production of and hydrogen by Chlorella vulgaris.

In our previous studies, Chlorella vulgaris had proven highly efficient in removing selenium (Se) from water, while the disposal of Se containing in algal biomass was still an issue of concern. Firstly, this research suggests algal Se could be released back to water, posing risks to aquatic wildlife. Thus, we further explored the possibility of using C.\xa0vulgaris to remove Se and produce and hydrogen simultaneously. Our results show the higher percentage of saturated fatty acids, especially , was found in the sulfur (S) deprived algae exposed to either selenate or selenite, although the highest content (21.9%) was found in the selenite treated algae in full BG11 medium. In addition, compared with the Se free algae, hydrogen production rate was 2.1- and 4.3-fold higher for the selenate and selenite treated algae, respectively. Se removal efficiency achieved by the selenite treated algae through accumulation and volatilization was 2.3-fold higher than the selenate treatment under hypoxic condition with S deprived, which is in contrast to the results obtained under aerobic conditions.Copyright © 2018. Published by Elsevier Ltd.

Keyword: fat metabolism

Mangiferin Improved Palmitate-Induced-Insulin Resistance by Promoting Free Fatty in HepG2 and C2C12 Cells via PPAR: Mangiferin Improved Insulin Resistance.

Elevated free fatty (FFA) is a key risk factor for insulin resistance (IR). Our previous studies found that mangiferin could decrease serum FFA levels in obese rats induced by a high- diet. Our research was to determine the effects and mechanism of mangiferin on improving IR by regulating FFA in HepG2 and C2C12 cells. The model was used to quantify PA-induced accumulation in the two cell lines treated with various concentrations of mangiferin simultaneously for 24\u2009h. We found that mangiferin significantly increased insulin-stimulated glucose uptake, via phosphorylation of protein kinase B (P-AKT), glucose transporter 2 (GLUT2), and glucose transporter 4 (GLUT4) protein expressions, and markedly decreased glucose content, respectively, in HepG2 and C2C12 cells induced by PA. Mangiferin significantly increased FFA uptake and decreased intracellular FFA and triglyceride (TG) accumulations. The activity of the peroxisome proliferator-activated receptor (PPAR) protein and its downstream proteins involved in fatty translocase (CD36) and carnitine palmitoyltransferase 1 (CPT1) and the fatty -oxidation rate corresponding to FFA were also markedly increased by mangiferin in HepG2 and C2C12 cells. Furthermore, the effects were reversed by siRNA-mediated knockdown of PPAR. Mangiferin ameliorated IR by increasing the consumption of glucose and promoting the FFA oxidation via the PPAR pathway in HepG2 and C2C12 cells.

Keyword: fat metabolism

Single Cell Oil Production from Undetoxified L. hydrolysate by .

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, . is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast from undetoxified lignocellulosic hydrolysates of steam-pretreated . The growth of was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total content was 64% of the dry cell weight and the yield was 63% of the theoretical. The fatty profile of triglycerides contained 27% , 33% oleic and 32% linoleic . These results proved the potential of production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Keyword: fat metabolism

Palm Oil and Beta-palmitate in Infant Formula: A Position Paper by the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) Committee on Nutrition.

Palm oil (PO) is used in infant formulas in order to achieve (PA) levels similar to those in human milk. PA in PO is esterified predominantly at the SN-1,3 position of triacylglycerol (TAG), and infant formulas are now available in which a greater proportion of PA is in the SN-2 position (typical configuration in human milk). As there are some concerns about the use of PO, we aimed to review literature on health effects of PO and SN-2-palmitate in infant formulas.PubMed and Cochrane Database of Systematic Reviews were systematically searched for relevant studies on possible beneficial effects or harms of either PO or SN-2-palmitate in infant formula on various health outcomes.We identified 12 relevant studies using PO and 21 studies using SN-2-palmitate. Published studies have variable methodology, subject characteristics, and some are underpowered for the key outcomes. PO is associated with harder stools and SN-2-palmitate use may lead to softer stool consistency. Bone effects seem to be short-lasting. For some outcomes (infant colic, faecal microbiota, ), the number of studies is very limited and summary evidence inconclusive. Growth of infants is not influenced. There are no studies published on the effect on markers of later diseases.There is insufficient evidence to suggest that PO should be avoided as a source of in infant formulas for health reasons. Inclusion of high SN-2-palmitate blend in infant formulas may have short-term effects on stool consistency but cannot be considered essential.

Keyword: fat metabolism

Feeding high-oleic peanuts to layer hens enhances egg yolk color and oleic fatty content in shell eggs.

Previous studies have identified normal-oleic peanuts as a suitable and economical broiler feed ingredient. However, no studies to date have examined the use of high-oleic (HO) peanut cultivars as a feed ingredient for laying hens and determined the impact of feeding HO peanuts on performance and egg nutritive qualities. This project aimed to examine the use of HO peanuts as a feed ingredient for layer hens to determine the effect on performance, egg chemistry, and quality of the eggs produced. Forty-eight 40-wk-old layer hens were fed a conventional soybean meal + corn control diet or a HO peanut + corn diet for 10 wk in conventional battery cages. Body and feed weights were collected weekly. Pooled egg samples were analyzed for quality, analysis, and peanut protein allergenicity. There were no significant differences in hen performance or egg quality as measured by USDA grade quality, egg albumen height, or egg Haugh unit between the treatment groups. However, eggs produced from layer hens fed the HO peanut + corn diet had reduced egg weights relative to the controls (P = 0.0001). Eggs produced from layer hens fed the HO peanut diet had greater yolk color scores (P < 0.0001), HO fatty (P < 0.0001), and β-carotene (P < 0.0001) levels in comparison to the controls. Eggs produced from hens fed the control diet had greater and stearic saturated fatty acids (P < 0.0001), and trans (P < 0.0001) content compared to eggs produced from hens fed the HO peanut diet. All egg protein extracts from all treatments at each time point were non-reactive with rabbit anti-peanut agglutinin antibodies. This study identifies HO peanuts as an abundant commodity that could be used to support local agricultural markets of peanuts and poultry within the southeastern United States and be of economic advantage to producers while providing a potential health benefit to the consumer with improved egg nutrition.Published by Oxford University Press on behalf of Poultry Science Association 2018.

Keyword: fat metabolism

Comparison of monoculture and mixed culture (Scenedesmus obliquus and wild algae) for C, N, and P removal and production.

This study compared the efficiency of nutrient removal and accumulation by a monoculture of Scenedesmus obliquus and mixed cultures of microalgae. The highest removal efficiencies of ammonium (99.2%), phosphate (91.2%), and total organic carbon (83.6%) occurred in the monoculture. All the mixed cultures were dominated by S. quadricauda; in some mixed cultures, the proportions of Chlamydomonas reinhardtii and C. microsphaera reached >\u200920%. The content and production in the monoculture were 15.9% and 52.3\xa0mg\xa0kg, respectively, significantly higher than those in all the mixed cultures of microalgae. In all the mixed cultures, the proportion of was >\u200950%. The results suggest that the monoculture had advantages over the mixed culture of microalgae in terms of nutrient removal and production.

Keyword: fat metabolism

Upregulation of miR-181a impairs by targeting PPARα expression in nonalcoholic fatty liver disease.

Recent studies have reported elevated expression of miR-181a in patients with non-alcoholic fatty liver disease (NAFLD), suggesting that it may play an important role in liver and insulin resistance. We aimed to investigate the effect of miR-181a in and find new treatments for NAFLD. The expression level of miR-181a in NAFLD patient serum and a (PA)-induced NAFLD cell model was examined by Q-PCR. Oil red O staining and triglyceride assays were used to assess accumulation in hepatocytes. Western blotting was used to detect the protein expression levels of peroxisome proliferator-activated receptor-α (PPARα) and the fatty β-oxidation-related genes. Direct interactions were validated by dual-luciferase reporter gene assays. MiR-181a expression was significantly upregulated in the serum of NAFLD patients and PA-induced hepatocytes. Inhibition of miR-181a expression resulted in the increased expression of PPARα and its downstream genes, and PA-induced accumulation in hepatocytes was also inhibited. Upregulation of miR-181a resulted in the downregulation of its direct target PPARα and downstream gene expression of PPARα as well as aggravated accumulation in hepatocytes. At the same time, the increased expression of PPARα can offset accumulation in hepatocytes induced by miR-181a mimics. This study demonstrates that reducing the expression of miR-181a may improve in NAFLD. The downregulation of miR-181a expression can be a therapeutic strategy for NAFLD by modulating its target PPARα.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Early and reversible changes to the hippocampal proteome in mice on a high- diet.

The rise in global obesity makes it crucial to understand how diet drives obesity-related health conditions, such as premature cognitive decline and Alzheimer\'s disease (AD). In AD hippocampal-dependent episodic memory is one of the first types of memory to be impaired. Previous studies have shown that in mice fed a high- diet (HFD) episodic memory is rapidly but reversibly impaired.In this study we use hippocampal proteomics to investigate the effects of HFD in the hippocampus. Mice were fed either a low- diet (LFD) or HFD containing either 10% or 60% (Kcal) from for 3\u2009days, 1\u2009week or 2\u2009weeks. One group of mice were fed the HFD for 1\u2009week and then returned to the LFD for a further week. Primary hippocampal cultures were challenged with (PA), the most common long-chain saturated FA in the Western diet, and with the anti-inflammatory, n-3 polyunsaturated FA, docosahexaenoic (DHA), or a combination of the two to ascertain effects of these fatty acids on dendritic structure.HFD-induced changes occur in hippocampal proteins involved in , inflammation, cell stress, cell signalling, and the cytoskeleton after 3\u2009days, 1\u2009week and 2\u2009weeks of HFD. Replacement of the HFD after 1\u2009week by a low- diet (LFD) for a further week resulted in partial recovery of the hippocampal proteome. Microtubule-associated protein 2 (MAP2), one of the earliest proteins changed, was used to investigate the impact of fatty acids (FAs) on hippocampal neuronal morphology. PA challenge resulted in shorter and less arborised dendrites while DHA had no effect when applied alone but counteracted the effects of PA when FAs were used in combination. Dendritic morphology recovered when PA was removed from the cell culture media.This study provides evidence for the rapid and reversible effects of diet on the hippocampal proteome and the impact of PA and DHA on dendritic structure.

Keyword: fat metabolism

Mst1 inhibition attenuates non-alcoholic fatty liver disease via reversing Parkin-related mitophagy.

Obesity-related non-alcoholic fatty liver disease (NAFLD) is connected with mitochondrial stress and hepatocyte apoptosis. Parkin-related mitophagy sustains mitochondrial homeostasis and hepatocyte viability. However, the contribution and regulatory mechanisms of Parkin-related mitophagy in NAFLD are incompletely understood. Macrophage stimulating 1 (Mst1) is a novel mitophagy upstream regulator which excerbates heart and cancer apoptosisn via repressing mitophagy activity. The aim of our study is to explore whether Mst1 contributes to NAFLD via disrupting Parkin-related mitophagy. A NAFLD model was generated in wild-type (WT) mice and Mst1 knockout (Mst1-KO) mice using high- diet (HFD). Cell experiments were conducted via (PA) treatment in the primary hepatocytes. The results in our study demonstrated that Mst1 was significantly upregulated in HFD-treated livers. Genetic ablation of Mst1 attenuated HFD-mediated hepatic injury and sustained hepatocyte viability. Functional studies illustrated that Mst1 knockdown reversed Parkin-related mitophagy and the latter protected mitochondria and hepatocytes against HFD challenge. Besides, we further figured out that Mst1 modulated Parkin expression via the AMPK pathway; blockade of AMPK repressed Parkin-related mitophagy and recalled hepatocytes mitochondrial apoptosis. Altogether, our data identified that NAFLD was closely associated with the defective Parkin-related mitophagy due to Mst1 upregulation. This finding may pave the road to new therapeutic modalities for the treatment of fatty liver disease.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Dietary habits affect fatty composition of visceral adipose tissue in subjects with colorectal cancer or obesity.

Aim of this study was to identify a possible relationship among dietary fatty acids (FA) intake, FA adipose tissue (AT) profile and cancer condition in lean vs obese subjects affected or not by colorectal cancer (CRC). Actually, inadequate dietary habits together with physical inactivity are primary determinants of obesity and cancer risk. Changes in play a crucial role in different types of cancer and key enzymes involved in -metabolic pathways, such as stearoyl-coA-desaturase 1 (SCD-1), are differentially expressed in normal and cancer tissues.Food frequency questionnaires (FFQ) were analyzed by Winfood software. FA were assessed by gas-liquid chromatography in visceral AT samples. Estimated desaturase activities were calculated as precursor FA/product FA ratio. Desaturase gene expressions were evaluated by RT-qPCR.Lean and obese CRC subjects showed inadequate dietary habits. In particular, lean CRC subjects showed increase in the intake of saturated FA, specifically (p\u2009=\u20090.0042) and stearic (p\u2009=\u20090.0091), and a corresponding reduction of monounsaturated FA consumption, in particular oleic (p\u2009=\u20090.002) with respect to lean without CRC. Estimated SCD-1 activity in AT was increased in all the groups vs lean without CRC (pANOVA\u2009=\u20090.029).Unhealthy eating habits, characterizing obese and CRC subjects, may influence the visceral AT profile and contribute to the alteration of the metabolic pathways. The quality of the diet, other than the quantity, can have a main role in the establishment of inflammatory microenvironment and in metabolic changes favouring CRC.

Keyword: fat metabolism

9-PAHSA promotes browning of white via activating G-protein-coupled receptor 120 and inhibiting lipopolysaccharide / NF-kappa B pathway.

Browning of white adipose tissue is a novel mechanism to counteract obesity in view of its thermogenic activity. Activation of G-protein-coupled receptor 120 (GPR120) can promote the browning of white . 9-PAHSA, an endogenous mammalian , which is acting as the ligand of GPR120 to enhance glucose uptake and exert anti-inflammatory effect. In the study, we would like to investigate the biological effects of 9-PAHSA on adipocyte browning. Here, we show that 9-PAHSA induces browning of 3T3-L1 adipocytes via enhanced expression of brown specific genes. 9-PAHSA-induced browning in white adipocytes of WT mice and ob/ob mice was investigated by determining expression levels of brown adipocyte-specific genes/proteins by quantitative real-time polymerase chain reaction analysis, immunoblot analysis and immunochemical staining. The effects of 9-PAHSA on brown markers in 3T3-L1 cells were decreased when GPR120 gene was silenced. To investigate the molecular mechanism of 9-PAHSA on adipocyte browning, lipopolysaccharide (LPS)-induced inflammatory model was conducted. 9-PAHSA treatment abolished LPS-induced NF-kappa B (NF-κB) activation and inflammatory cytokine secretion. But these anti-inflammatory effects of 9-PAHSA were attenuated by GPR120 knockdown. Our finding demonstrated that the browning of adipocyte was induced by 9-PAHSA through activating GPR120 and inhibiting the LPS/NF-κB pathway. This promising result will help to reveal the potential pathogenesis of obesity.Copyright © 2018. Published by Elsevier Inc.

Keyword: fat metabolism

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous fatty -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high- diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and . We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted fatty liver in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and fatty synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: fat metabolism

Aspalathin-Enriched Green Rooibos Extract Reduces Hepatic Insulin Resistance by Modulating PI3K/AKT and AMPK Pathways.

We previously demonstrated that an aspalathin-enriched green rooibos extract (GRE) reversed palmitate-induced insulin resistance in C2C12 skeletal muscle and 3T3-L1 cells by modulating key effectors of insulin signalling such as phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B (PI3K/AKT) and AMP-activated protein kinase (AMPK). However, the effect of GRE on hepatic insulin resistance is unknown. The effects of GRE on -induced hepatic insulin resistance using palmitate-exposed C3A liver cells and obese insulin resistant (OBIR) rats were explored. GRE attenuated the palmitate-induced impairment of glucose and in treated C3A cells and improved insulin sensitivity in OBIR rats. Mechanistically, GRE treatment significantly increased PI3K/AKT and AMPK phosphorylation while concurrently enhancing glucose transporter 2 expression. These findings were further supported by marked stimulation of genes involved in glucose , such as insulin receptor () and insulin receptor substrate 1 and 2 ( and ), as well as those involved in , including Forkhead box protein O1 (FOXO1) and carnitine palmitoyl transferase 1 (CPT1) following GRE treatment. GRE showed a strong potential to ameliorate hepatic insulin resistance by improving insulin sensitivity through the regulation of PI3K/AKT, FOXO1 and AMPK-mediated pathways.

Keyword: fat metabolism

Telmisartan protects against high glucose/high -induced apoptosis and insulin secretion by reducing the oxidative and ER stress.

Telmisartan, an angiotensin II receptor blocker, has been widely used for hypertension. It has also been reported to improve insulin sensitivity in animal models of obesity and diabetic patients by targeting to the peroxisome proliferator-activated receptor (PPAR)-γ. High glucose/high (HG/HL)-induced apoptosis of pancreatic β-cells impairs its function of insulin secretion and is generally believed to be the key factor in the development of diabetes. In this study, we investigated whether telmisartan exerted a protective effect against HG/HL-induced apoptosis and insulin secretion in vitro as well as in vivo; 10-μM telmisartan treatment significantly reduced HG (25\xa0mM) or/and HL (0.4\xa0mM ) induced-cell apoptosis and greatly improved insulin secretion in INS-1 pancreatic β-cells, which is consistent in an obesity rat model induced by HG/HL diets. Furthermore, telmisartan treatment markedly reduced the protein level of GRP78, CHOP, and caspase 12, while increasing anti-apoptotic Bcl-2 protein expression. Moreover, telmisartan treatment significantly reduced intracellular ROS levels. Mechanistically, we demonstrated that PPARγ signaling pathway may be involved in the telmisartan protective effects, which were blocked by a PPARγ blocker, GW9662. In conclusion, the protective effect of telmisartan was mediated by an anti-ER stress-induced apoptotic and anti-oxidative pathway. SIGNIFICANCE OF THIS STUDY: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder worldwide pathologically characterized by hyperglycemia and insulin resistance. Long-term high glucose in the blood has been proposed to induce pancreatic β-cell loss and is generally believed to be the key factor in the development of diabetes. In the present study, we demonstrated that telmisartan, a common drug used for hypertension treatment, has a protective effect against high glucose/high -induced cell apoptosis and greatly improves the insulin secretion function by inhibiting the oxidative stress and ER stress. Furthermore, this protective effect of telmisartan is mediated by the PPAR-γ signal pathway, which may provide a potential strategy against T2DM.© 2019 John Wiley & Sons, Ltd.

Keyword: fat metabolism

[Biological role of insulin: transformation of carnivores leaving in the ocean into herbivores living on the dry land. Development of the biological function of locomotion and kinetic parameters of the body.]

Phylogenetic theory of general pathology draws researchers\' attention to the following aspects of the biological effects of insulin. 1. Phylogenetically, in vivo insulin is primarily involved in the of fatty acids (FA) and only in the second turn in glucose ; regulation of FA in cells started millions of years earlier than that of glucose . Phylogenetically late insulin blocks lipolysis only in phylogenetically late insulin-dependent subcutaneous adipocytes but not in phylogenetically early visceral cells of the omentum. 2. Biological role of insulin consists in the formation of the biological function of locomotion, i.e., movement arising from contraction of striated myocytes provided with substrates (FA and glucose) for energy production as macroergic ATP. 3. Biological destiny of phylogenetically late insulin is transformation of carnivorous (fish-eating) animals living in the ocean into herbivores living on the dry land. 4. Insulin has formed in vivo highly efficient oleic variant of FA instead of phylogenetically early less efficient variant. 5. Biologically, insulin is destined for providing cells with energy and perfection of physical activity and kinetic parameters of the organism. According to phylogenetic theory of general pathology and basic principles of endocrinology, clinical cases should be regarded as: 1. rare structurally-related insulindeficient type I diabetes mellitus, 2. rare structurally-related (receptor pathology) hyperinsulinemic type II diabetes mellitus, and 3. a great number of patients with functional disorders in the hormone activity, insulin resistance syndrome, metabolic syndrome and obesity. We believe that the food of herbivores should contain in the amounts that hepatocytes can esterify into oleic very low density lipoproteins and cells can internalize as ligand oleic by apoЕ/В-100 endocytosis without formation of low density lipoproteins.

Keyword: fat metabolism

Resveratrol Ameliorates Droplet Accumulation in Liver Through a SIRT1/ ATF6-Dependent Mechanism.

droplets (LDs) are dynamic organelles that store neutral lipids during times of energy excess, and an increased accumulation of LDs in the liver is closely linked to hepatic steatosis. Our previous studies suggested that resveratrol (RSV) supplement could improve hepatic steatosis, but the underlying mechanism, particularly which related to LD accumulation, has not yet been elucidated.A high- diet (HFD) and were used to induce hepatic steatosis in mouse liver and hepatocytes, respectively. The effects of RSV on LD accumulation were analyzed in vivo and in vitro. The effects of RSV on the expression levels of LD-associated genes (ATF6, Fsp27β/CIDEC, CREBH, and PLIN1) were measured by qRT-PCR and western blot assays, followed by KD or overexpression of SIRT1 and ATF6 with small interfering RNAs or overexpressed plasmids, respectively. The dual luciferase reporter assay, chromatin immunoprecipitation assay, coimmunoprecipitation, and proximity ligation assay were utilized to clarify the mechanism of transcriptional regulation and possible interaction between SIRT1 and ATF6.There was a significant increase in the accumulation of LDs in liver and hepatocytes during the process of HFD-induced steatosis, respectively, which was significantly inhibited by RSV supplementation. RSV notably activated SIRT1 expression and decreased the expression levels of ATF6, Fsp27β/CIDEC, CREBH, and PLIN1, which are associated with LD accumulation. Interestingly, the inhibitory effects of RSV on LD accumulation and the associated expression of genes in hepatocytes were abrogated or strengthened with SIRT1 silencing or overexpression, respectively. On the contrary, the benefits of RSV in hepatocytes were eliminated or aggravated when transfected with the overexpressed ATF6 or ATF6 siRNA, respectively. Furthermore, we found that RSV stimulated SIRT1 expression significantly, which was followed by increased deacetylation and inactivation of ATF6, resulting in a positive feedback loop for SIRT1 transcription associated with ATF6 binding to the SIRT1 promoter region.Taken together, these findings indicate that RSV supplementation improves hepatic steatosis by ameliorating the accumulation of LDs, and this might be partially mediated by a SIRT1/ATF6-dependent mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fat metabolism

Relationship between acyl- and sterol metabolisms in diatoms.

Diatoms are a phylum of unicellular photosynthetic eukaryotes living in oceans and fresh waters, characterized by the complexity of their plastid, resulting from a secondary endosymbiosis event. In the model diatom Phaeodactylum tricornutum, fatty acids (FAs) are synthesized from acetyl-CoA in the stroma of the plastid, producing . FAs are elongated and desaturated to form very-long chain polyunsaturated fatty acids (VLC-PUFAs) in domains of the endomembrane system that need to be identified. Synthesis of VLC-PUFAs is coupled with their import to the core of the plastid via the so-called "omega" pathway. The biosynthesis of sterols in diatoms is likely to be localized in the endoplasmic reticulum as well as using precursors deriving from the mevalonate pathway, using acetyl-CoA as initial substrate. These metabolic modules can be characterized functionally by genetic analyzes or chemical treatments with appropriate inhibitors. Some \'metabolic modules\' are characterized by a very low level of metabolic intermediates. Since some chemical treatments or genetic perturbation of induce the accumulation of these intermediates, channeling processes are possibly involved, suggesting that protein-protein interactions might occur between enzymes within large size complexes or metabolons. At the junction of these modules, metabolic intermediates might therefore play dramatic roles in directing carbon fluxes from one direction to another. Here, acetyl-CoA seems determinant in the balance between TAGs and sterols. Future lines of research and potential utilization for biotechnological applications are discussed.Published by Elsevier B.V.

Keyword: fat metabolism

Fibroblast growth factor 21 protects against lipotoxicity-induced pancreatic β-cell dysfunction via regulation of AMPK signaling and .

Fibroblast growth factor 21 (FGF21) is known as a potent metabolic regulator but its protective mechanisms against lipotoxicity-induced β-cell dysfunction and apoptosis remain elusive. Here, we aimed to examine the regulatory pathways whereby FGF21 mediates islet in lipotoxicity-treated cells and animal models. Rat β-cell line (INS-1E cells) and islets isolated from C57/BL6J mice were exposed to (PA) with/without FGF21, mimicking lipotoxic conditions. Resultant insulin secretion and intracellular signaling were analyzed with Western blotting and RNA-seq. C57/BL6J and global FGF21 knockout (KO) mice were fed with a high- diet (HFD) to induce lipotoxicity and given with a long-acting mimetic of FGF21. Insulin resistance and β-cell function were then assessed using homeostasis model assessment of insulin resistance (HOMA-IR) and insulinogenic index. FGF21 ameliorated PA-induced accumulation, reversed cell apoptosis, and enhanced glucose-stimulated insulin secretion (GSIS) as impaired by lipotoxicity in islet β-cells. Mechanistically, FGF21 exerted its beneficial effects through activation of AMPK-ACC (acetyl-CoA carboxylase) pathway and peroxisome proliferation-activated receptors (PPARs) δ/γ signaling, thus increasing the levels of carnitine palmitoyltransferase-1A (CPT1A) and leading to increased fatty (FA) oxidation and reduced deposition in β-cells. Interestingly, FGF21 reduced PA-induced cell death via restoration of the expression of apoptosis inhibitor Birc3. In vivo studies further showed that FGF21 is critical for islet insulinogenic capacity and normal function in the context of HFD-treated animals. FGF21 down-regulates islet cell accumulation, probably via activation of AMPK-ACC and PPARδ/γ signaling, and reduces cell death under lipotoxicity, indicating that FGF21 is protective against lipotoxicity-induced β-cell dysfunction and apoptosis.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: fat metabolism

Factors associated with the content of mammary-synthesized fatty acids in milk : A meta-analysis.

Consumption of specific fatty acids (FA) that are synthesized in the mammary gland, namely de novo FA, has implications for human health. The objective of the present meta-analysis was to study the associations between milk content of de novo FA, with (1) diet composition, and (2) milk production and composition. Milk FA data from 96 peer-reviewed studies published between 1990 and 2016 that included 324 treatment means from 83 bovine experiments, 36 treatment means from 12 caprine experiments, and 40 treatment means from 12 ovine experiments were used in this analysis. Individual species models including the fixed effect of experiment were fitted using multiple regression to explain milk content of de novo FA as a function of diet composition and milk production and composition variables. We also evaluated replacing the effect of the experiment by the effect of the experiment nested in the laboratory at which the research had been conducted, and the effect of the laboratory. Butyric content in milk was positively but weakly related to dietary ether extract in does and ewes. Lauric, myristic, and contents in milk were negatively related to dietary ether extract in does and to a somewhat lesser extent in cows and ewes. The results confirm that the inclusion of lipids in the diet may not only affect the availability of preformed FA but also the profile of FA synthesized de novo in the mammary gland. Most of the variation in all prediction models was explained by the experiment or by the laboratory if the latter was included in the model. The ample variation in analytical methods reported by the different research groups suggests that differences in analytical protocols might explain a substantial proportion of the variation in de novo FA profile. A main conclusion of this study is the potential influence of differences in analytical procedures to explain the variation in de novo FA profile. Standardization of methods of FA analysis to improve reproducibility seems to be an aspect of importance to this area of research.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Flaxseed oil ameliorated high--diet-induced bone loss in rats by promoting osteoblastic function in rat primary osteoblasts.

α-Linolenic (ALA) is a plant-derived omega-3 unsaturated fatty that is rich in flaxseed oil (FO). The effect of FO on bone health is controversial. This study aims to evaluate the effect of FO on bone damage induced by a high- diet (HFD) and to explore the possible mechanism.Male Sprague-Dawley rats were fed a normal control diet (NC, 10% ), FO diet (NY, 10% ), HFD (60% ), or HFD containing 10% FO (HY, 60% ) for 22\u2009weeks. Micro CT and three-point bending tests were conducted to evaluate bone microstructure and biomechanics. Serum was collected for the detection of ALP, P1NP, and CTX-1. Rat primary osteoblasts (OBs) were treated with different concentrations of ALA with or without (PA) treatment. The ALP activity, osteogenic-related gene and protein expression were measured.Rats in the HFD group displayed decreased biomechanical properties, such as maximum load, maximum fracture load, ultimate tensile strength, stiffness, energy absorption, and elastic modulus, compared with the NC group ( \xa0<\u20090.05). However, HY attenuated the HFD-induced decreases in bone biomechanical properties, including maximum load, maximum fracture load, and ultimate tensile strength (\u2009<\u20090.05). Trabecular bone markers such as trabecular volume bone mineral density (Tb. vBMD trabecular bone volume/total volume (Tb. BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) were decreased, trabecular separation (Tb. Sp) and the structure model index (SMI) were increased in the HFD group compared with the NC group, and all parameters were remarkably improved in the HY group compared to the HFD group (p\u2009<\u20090.05). However, cortical bone markers such as cortical volume bone mineral density (Ct. vBMD), cortical bone volume/total volume (Ct. BV/TV) and cortical bone thickness (Ct. Th) were not significantly different among all groups. Moreover, the serum bone formation markers ALP and P1NP were higher and the bone resorption marker CTX-1 was lower in the HY group compared with levels in the HFD group. Compared with the NC group, the NY group had no difference in the above indicators. In rat primary OBs, PA treatment significantly decreased ALP activity and osteogenic gene and protein (β-catenin, RUNX2, and osterix) expression, and ALA dose-dependently restored the inhibition induced by PA.FO might be a potential therapeutic agent for HFD-induced bone loss, most likely by promoting osteogenesis.© The Author(s). 2019.

Keyword: fat metabolism

Anthocyanins ameliorate palmitate-induced inflammation and insulin resistance in 3T3-L1 adipocytes.

Increased adiposity has been associated with adipose tissue low-grade inflammation leading to insulin resistance. Adipocyte differentiation inhibitors are expected to be effective in preventing obesity and related diseases. Anthocyanins (ACNs) are associated to enhanced adipocyte function and protection from metabolic stress. Herein, we evaluated the in vitro protective effects of an ACN rich extract against (PA)-induced hypertrophy, inflammation, and insulin resistance in 3T3-L1 adipocytes. ACN extract pretreatment reduces accumulation and peroxisome proliferators-activated receptor-γ protein levels induced by PA. In addition, PA induces inflammation with activation of NF-κB pathway, whereas ACN extract pretreatment dose-dependently inhibited this pathway. Furthermore, adipocyte dysfunction associated with hypertrophy induces insulin resistance by affecting phosphatidylinositol 3-kinase-protein kinase B/Akt axis, GLUT-1, and adiponectin mRNA levels. ACN extract pretreatment reverts these effects induced by PA and moreover was able to induce insulin pathway with levels higher than insulin control cells, supporting an insulin sensitizer role for ACNs. This study demonstrates a prevention potential of ACNs against obesity comorbidities, due to their protective effects against inflammation/insulin resistance in adipocytes. In addition, these results contribute to the knowledge and strategies on the evaluation of the mechanism of action of ACNs from a food source under basal and insulin resistance conditions related to obesity.© 2019 John Wiley & Sons, Ltd.

Keyword: fat metabolism

1,25-Dihydroxyvitamin D regulates and glucose utilization in differentiated 3T3-L1 adipocytes.

It is well established that adipose tissue can both store and metabolize vitamin D. The active form of vitamin D, 1,25 dihydroxyvitamin D [1,25(OH)D], regulates adipocyte differentiation and inflammation, highlighting the multifaceted role that vitamin D plays in adipose tissue physiology. However, there is limited evidence regarding vitamin D regulation of mature adipocyte . We hypothesize that 1,25(OH)D alters and glucose in differentiated 3T3-L1 adipocytes to reduce triacylglycerol (TAG) accumulation. In this study, 1,25(OH)D (10\u202fnmol/L) stimulated a 21% reduction in TAG accumulation in differentiated 3T3-L1 adipocytes after 4\u202fdays (P\u202f=\u202f.01) despite a significant increase in fatty uptake (P\u202f<\u202f.01). Additionally, 1,25(OH)D stimulated a 2.5-fold increase in CO production from [1-C] (P\u202f<\u202f.01), indicative of an elevated rate of fatty β-oxidation, while stimulating a 9% reduction in de novo fatty synthesis (P\u202f=\u202f.03). Interestingly, d-[U-C]glucose incorporation into fatty acids was reduced by 30% in response to 1,25(OH)D (P\u202f<\u202f.01), indicating a reduced contribution of glucose as a substrate for de novo lipogenesis. Consistent with these findings, mRNA expression of the anaplerotic enzyme pyruvate carboxylase was reduced by 41% (P\u202f<\u202f.01). In summary, 1,25(OH)D stimulated fatty oxidation and reduced TAG accumulation in differentiated adipocytes. Furthermore, 1,25(OH)D reduced glucose utilization as a substrate for fatty synthesis potentially by downregulating pyruvate carboxylase and stimulating glucose disposal as glycerol. Collectively, these 1,25(OH)D-induced changes in and glucose utilization may contribute to the reduction in TAG accumulation and be protective against excessive mass accumulation and associated metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Neuroprotective effects of vitamin D on high diet- and -induced enteric neuronal loss in mice.

The role of vitamin D in obesity and diabetes is debated. Obese and/or diabetic patients have elevated levels of free fatty acids, increased susceptibility to gastrointestinal symptoms and are suggested to have altered vitamin D balance. The enteric nervous system is pivotal in regulating gastrointestinal activity and high diet (HFD) has been shown to cause loss of enteric neurons in ileum and colon. This study investigates the effect of vitamin D on HFD- and -induced enteric neuronal loss in vivo and in vitro.Mice were fed either a normal diet (ND) or HFD supplemented with varying levels of vitamin D (from 0x to 20x normal vitamin D level) for 19 weeks. Ileum and colon were analyzed for neuronal numbers and remodeling. Primary cultures of myenteric neurons from mouse small intestine were treated with (4x10M) and/or 1α,25-hydroxy-vitamin D3 (VD, 10- 10M) with or without modulators of and VD pathways. Cultures were analyzed by immunocyto- and histochemical methods.Vitamin D supplementation had no effect on enteric neuronal survival in the ND group. HFD caused substantial loss of myenteric neurons in ileum and colon. Vitamin D supplementation between 0-2x normal had no effect on HFD-induced neuronal loss. Supplementation with 20x normal, prevented the HFD-induced neuronal loss. In vitro supplementation of VD prevented the -induced neuronal loss. The VD receptor (VDR) was not identified in enteric neurons. Enteric glia expressed the alternative VD receptor, protein disulphide isomerase family A member 3 (PDIA3), but PDIA3 was not found to mediate the VD response in vitro. Inhibition of peroxisome proliferator-activated receptor gamma (PPARγ) and immune neutralization of isocitrate lyase prevented the VD mediated neuroprotection to exposure.Results show that VD protect enteric neurons against HFD and induced neuronal loss. The mechanism behind is suggested to be through activation of PPARγ leading to improved neuronal peroxisome function and of neuronal intermediates.

Keyword: fat metabolism

Protocatechuic -Ameliorated Endothelial Oxidative Stress through Regulating Acetylation Level via CD36/AMPK Pathway.

As one of the main metabolites of anthocyanin, protocatechuic (PCA) possesses strong antioxidant activity. In the present study, we explored the capacity of PCA on the alleviation of endothelial oxidative stress and investigated the underlying mechanisms using RNA sequencing (RNA-Seq). In comparison with (PA)-treated cells, PCA (100 μM) significantly decreased the generations of 3-nitrotyrosine (3-NT) and 8-hydroxydeoxyguanosine (8-OHdG) (0.82 ± 0.01 vs 1.16 ± 0.05 and 0.80 ± 0.01 vs 1.48 ± 0.15, respectively, p < 0.01), two biomarkers of oxidative damage, and restored the levels of nitric oxide (NO) (0.97 ± 0.04 vs 0.54 ± 0.02, p < 0.01) and mitochondrial membrane potential (MMP) (0.96 ± 0.03 vs 0.86 ± 0.02, p < 0.01) in human umbilical vein endothelial cells (HUVECs). PCA also obviously reduced the level of reactive oxygen species (ROS) (0.86 ± 0.15 vs 2.67 ± 0.09, p < 0.01) in aorta from high- diet (HFD)-fed mice. RNA-Seq and Western blot analysis indicated that PCA markedly reduced the expression of cluster of differentiation 36 (CD36), a membrane fatty transporter, and reduced the generations of adenosine triphosphate (ATP) and acetyl coenzyme A (Ac-CoA). These effects of PCA were associated with decreased level of acetylated-lysine and restored the activity of manganese-dependent superoxide dismutase (MnSOD) through reducing the generation of Ac-CoA or activating Sirt1 and Sirt3 via a CD36/AMP-kinase (AMPK) dependent pathway.

Keyword: fat metabolism

Concomitant production of chitosan and lipids from a newly isolated Mucor circinelloides ZSKP for biodiesel production.

A newly-isolated oleaginous fungus Mucor circinelloides ZSKP concurrently yielded 21.4% lipids and 11.2% chitosan per gram of biomass. Parameters affecting the co-production were identified using Plackett-Burman design and were statistically optimized using Response Surface Methodology, which resulted in a 3-fold improvement in production. The profile showed a high content of unsaturated fatty acids including oleic (37%), linolenic (14%) and linoleic acids (19%), while was the major saturated fatty (21%). A comparative study to evaluate the efficacy of enzymatic (lipase) and chemical treatments for biodiesel production from fungal lipids and sunflower oil revealed enhanced production of biodiesel from fungal lipids. Synthesized biodiesel from M. circinelloides ZSKP satisfied standard specifications and had a higher cetane number (56), lower kinematic viscosity (4.6\u202fmm/s) and lower number (0.03) compared to sunflower oil. Results suggest Mucor circinelloides ZSKP is a promising candidate for implementation of the biorefinery concept.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Nutrient-Dependent Changes of Protein Palmitoylation: Impact on Nuclear Enzymes and Regulation of Gene Expression.

Diet is the main environmental stimulus chronically impinging on the organism throughout the entire life. Nutrients impact cells via a plethora of mechanisms including the regulation of both protein post-translational modifications and gene expression. Palmitoylation is the most-studied protein lipidation, which consists of the attachment of a molecule of to residues of proteins. -palmitoylation is a reversible cysteine modification finely regulated by palmitoyl-transferases and acyl-thioesterases that is involved in the regulation of protein trafficking and activity. Recently, several studies have demonstrated that diet-dependent molecules such as insulin and fatty acids may affect protein palmitoylation. Here, we examine the role of protein palmitoylation on the regulation of gene expression focusing on the impact of this modification on the activity of chromatin remodeler enzymes, transcription factors, and nuclear proteins. We also discuss how this physiological phenomenon may represent a pivotal mechanism underlying the impact of diet and nutrient-dependent signals on human diseases.

Keyword: fat metabolism

GPR120 protects lipotoxicity-induced pancreatic β-cell dysfunction through regulation of PDX1 expression and inhibition of islet inflammation.

G-protein coupled receptor 120 (GPR120) has been shown to act as an omega-3 unsaturated fatty sensor and is involved in insulin secretion. However, the underlying mechanism in pancreatic β cells remains unclear. To explore the potential link between GPR120 and β-cell function, its agonists docosahexaenoic (DHA) and GSK137647A were used in (PA)-induced pancreatic β-cell dysfunction, coupled with GPR120 knockdown (KD) in MIN6 cells and GPR120 knockout (KO) mice to identify the underlying signaling pathways. and treatments of MIN6 cells and islets isolated from wild-type (WT) mice with DHA and GSK137647A restored pancreatic duodenal homeobox-1 (PDX1) expression levels and β-cell function via inhibiting PA-induced elevation of proinflammatory chemokines and activation of nuclear factor κB, c-Jun amino (N)-terminal kinases1/2 and p38MAPK signaling pathways. On the contrary, these GPR120 agonism-mediated protective effects were abolished in GPR120 KD cells and islets isolated from GPR120 KO mice. Furthermore, GPR120 KO mice displayed glucose intolerance and insulin resistance relative to WT littermates, and β-cell functional related genes were decreased while inflammation was exacerbated in islets with increased macrophages in pancreas from GPR120 KO mice. DHA and GSK137647A supplementation ameliorated glucose tolerance and insulin sensitivity, as well as improved expression and islet inflammation in diet-induced obese WT mice, but not in GPR120 KO mice. These findings indicate that GPR120 activation is protective against lipotoxicity-induced pancreatic β-cell dysfunction, via the mediation of PDX1 expression and inhibition of islet inflammation, and that GPR120 activation may serve as a preventative and therapeutic target for obesity and diabetes.© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: fat metabolism

Effect of Microalgae Polysaccharides on Biochemical and Metabolomics Pathways Related to Plant Defense in Solanum lycopersicum.

Microalgae are photosynthetic microorganisms that produce several bioactive molecules that have received considerable attention in scientific and industrial communities. Today, many plant biostimulants including seaweed extracts and polysaccharides are used in agriculture. However, microalgae have not been largely exploited in this field as a potential source of plant bio stimulants. This study investigated the biostimulatory effects of microalgae polysaccharides on different metabolomic and biochemical pathways related to plant defense. 0.2\xa0mg\xa0mL of crude polysaccharides extracted from four green microalgae strains was injected into tomato plants (Solanum lycopersicum). β-1,3-glucanase activity, remodeling, phenylalanine ammonia lyase (PAL), Lipoxygenase (LOX), and antioxidant enzyme (APX, POD and CAT) activities were evaluated 48\xa0h after treatment. Plants treated with crude polysaccharides extracted from. C. vulgaris and C. sorokiniana exhibited a significant increase in β-1,3-glucanase activity. Accordingly, C. sorokiniana crude polysaccharides had a significant stimulatory effect on PAL activity with a percentage increase of 188.73% compared to the control. GC/MS quantitative lipidomics analysis revealed that treatment with D. salina, C. sorokiniana, and C. reinhardtii crude polysaccharides increased PUFA content by 50.37%, 34.46%, and 33.37% respectively. Microalgae polysaccharides also enhanced stearic , , and VLCFA content, the optimal value of which increased by 45.50%, 32.83%, and 60.60% respectively under treatment with C. reinhardtii crude polysaccharides compared with the control. C. vulgaris and C. reinhardtii crude polysaccharides also exhibited higher APX and POD activity respectively. The present results therefore indicate the potentiality of microalgae crude polysaccharides as a promising renewable bio resource in the development plant bio stimulants.

Keyword: fat metabolism

Saturated fatty stimulates production of extracellular vesicles by renal tubular epithelial cells.

Lipotoxicity, an accumulation of intracellular metabolites, has been proposed as an important pathogenic mechanism contributing to kidney dysfunction in the context of metabolic disease. , a predominant derivative, can cause lipoapoptosis and the release of inflammatory extracellular vesicles (EVs) in hepatocytes, but the effect of lipids on EV production in chronic kidney disease remains vaguely explored. This study was aimed to investigate whether would stimulate EV release from renal proximal tubular epithelial cells. Human and rat proximal tubular epithelial cells, HK-2 and NRK-52E, were incubated with 1% bovine serum albumin (BSA), BSA-conjugated (PA), and BSA-conjugated oleic (OA) for 24-48\xa0h. The EVs released into conditioned media were isolated by ultracentrifugation and quantified by nanoparticle-tracking analysis (NTA). According to NTA, the size distribution of EVs was 30-150\xa0nm with similar mode sizes in all experimental groups. Moreover, BSA-induced EV release was significantly enhanced in the presence of PA, whereas EV release was not altered by the addition of OA. In NRK-52E cells, PA-enhanced EV release was associated with an induction of cell apoptosis reflected by an increase in cleaved caspase-3 protein by Western blot and Annexin V positive cells analyzed by flow cytometry. Additionally, confocal microscopy confirmed the uptake of -induced EVs by recipient renal proximal tubular cells. Collectively, our results indicate that PA stimulates EV release from cultured proximal tubular epithelial cells. Thus, extended characterization of -induced EVs may constitute new signaling paradigms contributing to chronic kidney disease pathology.

Keyword: fat metabolism

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced insulin sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe insulin antagonism and body mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived fatty acids may impair insulin sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum fatty liver disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated -induced insulin resistance that defines in part the mechanisms of type 2 diabetes mellitus and nonalcoholic fatty liver disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of insulin-stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit insulin action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and fatty , insulin function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Sodium Orthovanadate Changes Fatty Composition and Increased Expression of Stearoyl-Coenzyme A Desaturase in THP-1 Macrophages.

Vanadium compounds are promising antidiabetic agents. In addition to regulating glucose , they also alter . Due to the clear association between diabetes and atherosclerosis, the purpose of the present study was to assess the effect of sodium orthovanadate on the amount of individual fatty acids and the expression of stearoyl-coenzyme A desaturase (SCD or Δ-desaturase), Δ-desaturase, and Δ-desaturase in macrophages. THP-1 macrophages differentiated with phorbol 12-myristate 13-acetate (PMA) were incubated in vitro for 48\xa0h with 1\xa0μM or 10\xa0μM sodium orthovanadate (NaVO). The estimation of fatty composition was performed by gas chromatography. Expressions of the genes SCD, fatty desaturase 1 (FADS1), and fatty desaturase 2 (FADS2) were tested by qRT-PCR. Sodium orthovanadate in THP-1 macrophages increased the amount of saturated fatty acids (SFA) such as and stearic , as well as monounsaturated fatty acids (MUFA)-oleic and palmitoleic . Sodium orthovanadate caused an upregulation of SCD expression. Sodium orthovanadate at the given concentrations did not affect the amount of polyunsaturated fatty acids (PUFA) such as linoleic , arachidonic , eicosapentaenoic (EPA), docosapentaenoic (DPA), and docosahexaenoic (DHA). In conclusion, sodium orthovanadate changed SFA and MUFA composition in THP-1 macrophages and increased expression of SCD. Sodium orthovanadate did not affect the amount of any PUFA. This was associated with a lack of influence on the expression of FADS1 and FADS2.

Keyword: fat metabolism

Plasma fatty acids in de novo lipogenesis pathway are associated with diabetogenic indicators among adults: NHANES 2003-2004.

Insulin regulates fatty acids (FAs) in the blood; conversely, FAs may mediate insulin sensitivity and are potentially modifiable risk factors of the diabetogenic state.The objective of our study was to examine the associations between plasma concentrations of FAs, fasting plasma glucose (FPG), and glycated hemoglobin (HbA1c) among individuals (n\xa0=\xa01433) in the NHANES (2003-2004).Plasma concentrations of 24 individual FAs were considered individually and in subgroups, per chemical structure. Study participants were categorized in diabetogenic groups: Group 1 (HbA1c ≥6.5% or FPG ≥126 mg/dL), Group 2 (HbA1c 5.7% to <6.5% or FPG 100 to <126 mg/dL), and Group 3 (HbA1c <5.7% and FPG <100 mg/dL). We assessed associations between diabetogenic groups and plasma FAs in multivariate multinomial regressions (with Group 3 as the reference).Overall, 7.0% of study participants were in Group 1; 33.3% were in Group 2. Plasma concentrations of several individual FAs, including even-chain saturated FAs (SFAs; myristic, , stearic acids) and monounsaturated FAs (MUFAs; cis-vaccenic, oleic acids), were respectively associated with greater odds of Groups 1 and 2 status, adjusting for covariates. Higher concentrations of SFA and MUFA subgroups (highest compared with lowest quartile) were associated with increased odds of Group 2 status [SFAs adjusted OR (aOR): 1.51 (95% CI: 1.05, 2.18); MUFAs aOR: 1.78 (95% CI: 1.11, 2.85)]. Higher eicosapentaenoic plasma concentration was associated with decreased odds of Group 1 status [quartile 4 aOR: 0.41 (95% CI: 0.17, 0.95)].Higher plasma concentrations of SFAs and MUFAs, primary de novo lipogenesis products, were associated with elevated FPG and HbA1c in a nationally representative study population in the United States. Additional studies are necessary to elucidate potential causal relationships between FAs (from endogenous production and dietary consumption) and diabetogenic indicators, as well as clinical implications for managing diabetes and prediabetes.

Keyword: fat metabolism

PKCζ Phosphorylates SIRT6 to Mediate Fatty β-Oxidation in Colon Cancer Cells.

Protein kinase C (PKC) has critical roles in regulating anabolism and catabolism. PKCζ, a member of atypical PKC family, has been reported to mediate glucose . However, whether and how PKCζ regulates tumor cells fatty β-oxidation are unknown. Here, we report that the phosphorylation of SIRT6 is significantly increased after (PA) treatment in colon cancer cells. PKCζ can physically interact with SIRT6 in vitro and in vivo, and this interaction enhances following PA treatment. Further experiments show that PKCζ is the phosphorylase of SIRT6 and phosphorylates SIRT6 at threonine 294 residue to promote SIRT6 enrichment on chromatin. In the functional study, we find that the expression of ACSL1, CPT1, CACT, and HADHB, the genes related to fatty β-oxidation, increases after PA stimulation. We further confirm that PKCζ mediates the binding of SIRT6 specifically to the promoters of fatty β-oxidation-related genes and elicits the expression of these genes through SIRT6 phosphorylation. Our findings demonstrate the mechanism of PKCζ as a new phosphorylase of SIRT6 on maintaining tumor fatty β-oxidation and define the new role of PKCζ in homeostasis.Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

[Lipolysis in very low density lipoproteins - locus minoris resistentiae - in the pathogenesis of hypertriglyceridemia. Positive effects of diet, polyenic fatty acids, statins and fibrates.]

Inhibition of hydrolysis of and oleic triglycedires (TG) in very low density lipoproteins (VLDL), slow formation of active apoВ-100 conformation, blockade of апоЕ/В-100 ligand formation in VLDL and their reduced uptake by insulin-dependent cells cause hypertriglyceridemia (HTG). and oleic VLDL (>80% total VLDL) are not converted in low density lipoproteins (LDL). Atherosclerosis is not an alimentary deficiency of polyenic fatty acids (PFA), but results from low in vivo bioavailability of PFA in LDL against the background of high dietary FA and LDL. Plasma PFA content and cellular PFA deficiency are as high as LDL cholesterol (CL). Primary prevention of atherosclerosis should be based on a decrease in dietary content of saturated FA, trans FA and a moderate increase in PFA. It seems highly unlikely that the xeobiotics statins, fibrates and probucol produce pleiotropic biological effects in vivo. These effects are brought about by phylogenetically early humoral mediators eicosanoids: prostacyclins, prostaglandins, thromboxanes, leukotrienes, and resolvins. It is reasonable to suggest that all preparations which act according to the same algorithm activate TG hydrolysis in VLDL and normalize cellular uptake of PFA in linoleic and linolenic LDL via apoВ-100 endocytosis. Atherosclerosis is a syndrome of cellular deficiency of essential polyenic FA.

Keyword: fat metabolism

Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Fatty Liver Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high- diet (HFD)-induced non-alcoholic fatty liver disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified fatty levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic lipogenesis-related gene expression, sterol regulatory element-binding transcription protein 1 (, fatty synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the liver, and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated accumulation and macrophage-mediated inflammation.

Keyword: fat metabolism

Rosiglitazone ameliorates -induced cytotoxicity in TM4 Sertoli cells.

The Sertoli cell is the only somatic cell within the seminiferous tubules, and is vital for testis development and spermatogenesis. Rosiglitazone (RSG) is a member of the thiazolidinedione family and is a peroxisome proliferator-activated receptor-γ (PPARγ) agonist. It has been reported that RSG protects various types of cells from fatty -induced damage. However, whether RSG serves a protective role in Sertoli cells against (PA)-induced toxicity remains to be elucidated. Therefore, the aim of the present study was to investigate the effect of RSG on PA-induced cytotoxicity in Sertoli cells. MTT assay and Oil Red O staining revealed that RSG ameliorated the PA-induced decrease in TM4 cell viability, which was accompanied by an alleviation of PA-induced accumulation in cells. In primary mouse Sertoli cells, RSG also showed similar protective effects against PA-induced lipotoxicity. Knockdown of PPARγ verified that RSG exerted its protective role in TM4 cells through a PPARγ-dependent pathway. To evaluate the mechanism underlying the protective role of RSG on PA-induced lipotoxicity, the present study analyzed the effects of RSG on PA uptake, and the expression of genes associated with both fatty oxidation and triglyceride synthesis. The results demonstrated that although RSG did not affect the endocytosis of PA, it significantly elevated the expression of carnitine palmitoyltransferase (CPT)-1A, a key enzyme involved in fatty oxidation, which indicated that the protective effect of RSG may have an important role in fatty oxidation. On the other hand, the expression of CPT1B was not affected by RSG. Moreover, the expression levels of diacylglycerol O-acyltransferase (DGAT)-1 and DGAT2, both of which encode enzymes catalyzing the synthesis of triglycerides, were not suppressed by RSG. The results indicated that RSG reduced PA-induced accumulation by promoting fatty oxidation mediated by CPT1A. The effect of RSG in protecting cells from lipotoxicity was also found to be specific to Sertoli cells and hepatocytes, and not to other cell types that do not store excess in large quantities, such as human umbilical vein endothelial cells. These findings provide insights into the cytoprotective effects of RSG on Sertoli cells and suggest that PPARγ activation may be a useful therapeutic method for the treatment of Sertoli cell dysfunction caused by dyslipidemia.

Keyword: fat metabolism

Increased triacylglycerol - Fatty substrate cycling in human skeletal muscle cells exposed to eicosapentaenoic .

It has previously been shown that pretreatment of differentiated human skeletal muscle cells (myotubes) with eicosapentaenoic (EPA) promoted increased uptake of fatty acids and increased triacylglycerol accumulation, compared to pretreatment with oleic (OA) and (PA). The aim of the present study was to examine whether EPA could affect substrate cycling in human skeletal muscle cells by altering lipolysis rate of intracellular TAG and re-esterification of fatty acids. Fatty was studied in human myotubes using a mixture of fatty acids, consisting of radiolabelled oleic as tracer (14C-OA) together with EPA or PA. Co-incubation of myotubes with EPA increased cell-accumulation and incomplete fatty oxidation of 14C-OA compared to co-incubation with PA. distribution showed higher incorporation of 14C-OA into all cellular lipids after co-incubation with EPA relative to PA, with most markedly increases (3 to 4-fold) for diacylglycerol and triacylglycerol. Further, the increases in cellular lipids after co-incubation with EPA were accompanied by higher lipolysis and fatty re-esterification rate. Correspondingly, basal respiration, proton leak and maximal respiration were significantly increased in cells exposed to EPA compared to PA. Microarray and Gene Ontology (GO) enrichment analysis showed that EPA, related to PA, significantly changed i.e. the GO terms "Neutral metabolic process" and "Regulation of storage". Finally, an inhibitor of diacylglycerol acyltransferase 1 decreased the effect of EPA to promote fatty accumulation. In conclusion, incubation of human myotubes with EPA, compared to PA, increased processes of fatty turnover and oxidation suggesting that EPA may activate futile substrate cycling of fatty acids in human myotubes. Increased TAG-FA cycling may be involved in the potentially favourable effects of long-chain polyunsaturated n-3 fatty acids on skeletal muscle and whole-body energy .

Keyword: fat metabolism

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic fatty liver disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high- diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and liver-to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and -uptake markers and less signs of inflammation in the liver. GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human liver biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by fatty acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of fatty (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA into herbivorous dry land-living species with oleic type of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic fatty liver disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: fat metabolism

Evidence for perturbed metabolic patterns in bipolar disorder subjects associated with lithium responsiveness.

Bipolar disorder (BD) is multifactorial mood disorder characterized by alternating episodes of hyperactive mania and severe depression. Lithium is one of the most preferred drug used as mood stabilizer in treating BD. In this study, we examined the changes in plasma metabolome in BD subjects in the context of lithium responsiveness. Plasma samples from clinically defined, age and gender matched unrelated healthy controls and BD subjects (lithium responders and non-responders) were obtained and processed in positive and negative mode using untargeted liquid chromatography/mass spectrometry analysis. We identified significant alterations in plasma levels of dopamine along with its precursors (tyrosine and phenylalanine), branched chain amino such as valine and excitatory neurotransmitter glutamate between healthy control and BD subjects. molecules such as, eicosenoic and retinyl ester also showed distinguished patterns between control and BD individuals. Lithium responsiveness was markedly associated with significant differences in proline, L-gamma-glutamyl-isoleucine, dopamine, methyl ester, cholesterol sulfate, androsterone sulfate and 9S,12S,13S-triHOME levels. Altered metabolites enriched with key biochemical pathways associated with neuropsychiatry disorders. We hypothesize that BD pathogenesis and lithium responsiveness is associated with impaired homeostasis of amino and .Copyright © 2019. Published by Elsevier B.V.

Keyword: fat metabolism

Ellagic Suppresses the Oxidative Stress Induced by Dietary-Oxidized Tallow.

Dietary tallow was thermally oxidized at 180°C in an open fryer. The oxidized tallow (OT) and unoxidized tallow were characterized for oxidation parameters and fatty composition using GC-MS. Tallow samples were fed to rabbits along with 50, 100, and 150\u2009mg/kg/day of ellagic (EA) for three weeks. Results revealed that the peroxide value (PV) and thiobarbituric reactive substances (TBARS) significantly increased, while radical scavenging activity (RSA) of the tallow decreased significantly with oxidation. GC-MS analysis showed eight fatty acids in the tallow samples, where (48.5-49.7\u2009g/100\u2009g), linoleic (18.7-23.7\u2009g/100\u2009g), stearic (13.5-15.6\u2009g/100\u2009g), and margaric (6.32-6.42\u2009g/100\u2009g) were the major fatty acids. Animal studies showed that oxidized tallow (OT) alone or in combination with EA significantly altered the body weight of the rabbits. Serum biochemical parameters and renal function tests were affected by OT and ameliorated by EA. The toxic effects of OT on haematological indices were minimized by EA. The supplementation of OT alone had significant effects on the liver structure and functions. The coadministration of EA reduced the toxic properties of OT on the liver, by increasing the antioxidant (GSH) system. The rabbit heart was also affected by the OT, which was ameliorated by EA supplementation. These results suggested that the supplementation of EA was beneficial against the OT-induced oxidative stress and may be considered for foods containing oxidized lipids.

Keyword: fat metabolism

The effect of enterolactone on liver precursors of inflammation.

The aim of this study was to assess the effects of enterolactone (ENL) on fractions fatty acids composition affecting hepatocyte inflammation development.The experiments were conducted in HepG2 cells incubated with ENL and/or (16\u202fh). Intracellular contents of free fatty acids (FFA), di- (DAG) and tri- (TAG) acylglycerol as well as their fatty acids compositions were assessed by Gas-Liquid Chromatography. Moreover, the ω-6/ω-3 ratios in the above mentioned lipids fractions were estimated. The expression of proteins involved in eicosanoids and prostanoids production (COX-2, 15-LOX), inflammatory process (TNFα), as well as the proteins participating in the desaturation (SCD 1) and elongation (Elovl 3, Elovl 6) of fatty acids were evaluated by Western Blot.Enterolactone modified fatty acids composition in FFA, DAG and TAG fractions. In conjunction with overload, it increased the content of ω-6 more than ω-3 PUFA. Moreover, it enhanced the expressions of Elovl 3, Elovl 6, COX-2 and TNFα, whereas it had no influence on SCD 1 and 15-LOX level.Our study revealed that the supplementation with ENL affected intracellular hepatic composition of saturated as well as unsaturated fatty acids in each of the investigated fractions. Based on the shift in the ω-6/ω-3 balance towards ω-6, as well as the increase in COX-2 and TNFα protein expressions, we may postulate a pro-inflammatory nature of the examined polyphenol. Moreover, our findings could prove to be useful in the future research in the topic of widespread diseases such as NASH.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Molecular mechanism of activation of the immunoregulatory amidase NAAA.

Palmitoylethanolamide is a bioactive that strongly alleviates pain and inflammation in animal models and in humans. Its signaling activity is terminated through degradation by -acylethanolamine amidase (NAAA), a cysteine hydrolase expressed at high levels in immune cells. Pharmacological inhibitors of NAAA activity exert profound analgesic and antiinflammatory effects in rodent models, pointing to this protein as a potential target for therapeutic drug discovery. To facilitate these efforts and to better understand the molecular mechanism of action of NAAA, we determined crystal structures of this enzyme in various activation states and in complex with several ligands, including both a covalent and a reversible inhibitor. Self-proteolysis exposes the otherwise buried active site of NAAA to allow catalysis. Formation of a stable substrate- or inhibitor-binding site appears to be conformationally coupled to the interaction of a pair of hydrophobic helices in the enzyme with membranes, resulting in the creation of a linear hydrophobic cavity near the active site that accommodates the ligand\'s acyl chain.

Keyword: fat metabolism

Rare ginsenosides ameliorate overload-induced myocardial insulin resistance via modulating metabolic flexibility.

Rare ginsenosides are found in ginseng and notoginseng, two medicinal plants widely used in China for treatment of cardiovascular diseases and type 2 diabetes. However, their pharmacological studies regarding myocardial fuel and insulin signaling are not clear.To explore the effect of a rare ginsenoside-standardized extract (RGSE), derived from steamed notoginseng, on cardiac fuel and insulin signaling.We used (PA) to treat H9c2 cells in vitro and high diet (HFD) to mice to induce insulin resistance in vivo.In vitro, differentiated H9c2 cells were pretreated with RGSE, metformin, mildronate or dichloroacetate (DCA) and stimulated with PA. In vivo, mice were fed with HFD and received RGSE, metformin or DCA for 6 weeks. Protein expression was determined by Western blotting. Mitochondrial membrane potential (Δψm), glucose uptake and reactive oxygen species (ROS) production were measured by fluorescence labeling. Other assessments including oxygen consumption rate (OCR) were also performed.RGSE prevented PA-induced decrease in pyruvate dehydrogenase (PDH) activity and increase in carnitine palmitoyltransferase 1 (CPT1) expression, and ameliorated insulin-mediated glucose uptake and utilization in H9c2 cells. Metformin and mildronate exhibited similar effects. In vivo, RGSE counteracted HFD-induced increase in myocardial expression of p-PDH and CPT1 and ameliorated cardiac insulin signaling. Metformin and DCA also showed beneficial effects. Further study showed that RGSE decreased OCR and mitochondrial complex I activity in PA-treated H9c2 cells, reduced ROS production and relieved mitochondrial oxidative stress, thus decreased serine phosphorylation in IRS-1.RGSE ameliorated myocardial insulin sensitivity under conditions of overload, which was tightly associated with the decrease in mitochondrial oxidative stress via modulating glucose and fatty oxidation.Copyright © 2018. Published by Elsevier GmbH.

Keyword: fat metabolism

Influence of Resveratrol on Sphingolipid in Hepatocellular Carcinoma Cells in Overload State.

Obesity is characterized by increased long chain fatty acids (LCFA) uptake and impaired in hepatocytes. Consequently, an enhanced intracellular content, including sphingolipids, may lead to lipotoxicity. It is believed that resveratrol (RSV), one of the most extensively studied plant-derived polyphenols, and its interaction with sphingolipid may constitute one of the major therapeutic targets for cancer and metabolic diseases treatment.The aim of this study was to ascertain, whether resveratrol may affect sphingolipid metabolic pathways, enzymes and transporters in a overload state.The experiments were conducted on hepatocellular carcinoma cells (HepG2) incubated with RSV and/or (PA) at the concentration of 0.5 mM and 50 µM, respectively for 16h. Intra- and extracellular sphingolipid concentrations were assessed by high-performance liquid chromatography and gas liquid chromatography. Moreover, the expression of caspase 3, selected fatty transporters and sphingolipid pathway proteins were estimated by Western Blot.RSV alone and together with PA significantly increased the intracellular concentration of ceramide, sphinganine and sphingosine as well as the expression of enzymes related to de novo ceramide synthesis pathway. Moreover, in our study, we observed augmented ceramide and sphingomyelin efflux into the incubation media in these groups. In addition, RSV substantially reduced intracellular triacylglycerols accumulation in overload conditions.The above-mentioned findings suggest that RSV, at least partially, demonstrates a potential protective effect on HepG2 cells in a overload state.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: fat metabolism

Obesity-induced activation of JunD promotes myocardial accumulation and metabolic cardiomyopathy.

Metabolic cardiomyopathy (MC)-characterized by intra-myocardial triglyceride (TG) accumulation and lipotoxic damage-is an emerging cause of heart failure in obese patients. Yet, its mechanisms remain poorly understood. The Activator Protein 1 (AP-1) member JunD was recently identified as a key modulator of hepatic in obese mice. The present study investigates the role of JunD in obesity-induced MC.JunD transcriptional activity was increased in hearts from diet-induced obese (DIO) mice and was associated with myocardial TG accumulation and left ventricular (LV) dysfunction. Obese mice lacking JunD were protected against MC. In DIO hearts, JunD directly binds PPARγ promoter thus enabling transcription of genes involved in TG synthesis, uptake, hydrolysis, and storage (i.e. Fas, Cd36, Lpl, Plin5). Cardiac-specific overexpression of JunD in lean mice led to PPARγ activation, cardiac steatosis, and dysfunction, thereby mimicking the MC phenotype. In DIO hearts as well as in neonatal rat ventricular myocytes exposed to , Ago2 immunoprecipitation, and luciferase assays revealed JunD as a direct target of miR-494-3p. Indeed, miR-494-3p was down-regulated in hearts from obese mice, while its overexpression prevented lipotoxic damage by suppressing JunD/PPARγ signalling. JunD and miR-494-3p were also dysregulated in myocardial specimens from obese patients as compared with non-obese controls, and correlated with myocardial TG content, expression of PPARγ-dependent genes, and echocardiographic indices of LV dysfunction.miR-494-3p/JunD is a novel molecular axis involved in obesity-related MC. These results pave the way for approaches to prevent or treat LV dysfunction in obese patients.Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.

Keyword: fat metabolism

A Pathophysiological Model of Non-Alcoholic Fatty Liver Disease Using Precision-Cut Liver Slices.

Non-alcoholic fatty liver disease (NAFLD) is a common liver disorder closely related to metabolic syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between liver cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut liver slices (PCLSs) were used in this study. PCLSs, prepared from liver tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic metabolic syndrome. Accumulation of droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired fatty transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: fat metabolism

Chitosan Oligosaccharides Improve Glucolipid Disorder in Liver by Suppression of Obesity-Related Inflammation and Restoration of Peroxisome Proliferator-Activated Receptor Gamma (PPARγ).

Chitosan oligosaccharides (COS) display various biological activities. In this study, we aimed to explore the preventive effects of COS on glucolipid disorder using (PA)-induced HepG2 cells and high- diet (HFD)-fed C57BL/6J mice as experimental models in vitro and in vivo, respectively. The results showed that COS pretreatment for 12 h significantly ameliorated accumulation in HepG2 cells exposed to PA for 24 h, accompanied by a reversing of the upregulated mRNA expression of proinflammatory cytokines (IL-6, MCP-1, TNF-α) and glucolipid -related regulators (SCD-1, ACC1, PCK1-α). In addition, COS treatment alleviated glucolipid disorder in mice fed with HFD for five months, including reduction in body weight and fasting glucose, restoration of intraperitoneal glucose tolerance, and suppression of overexpression of proinflammatory cytokines and glucolipid -related regulators. Furthermore, our study found that COS pretreatment significantly reversed the downregulation of PPARγ at transcriptional and translational levels in both PA-induced HepG2 cells and liver tissues of HFD-fed mice. In summary, the study suggests that COS can improve glucolipid disorder by suppressing inflammation and upregulating PPARγ expression. This indicates a novel application of COS in preventing and treating glucolipid -related diseases.

Keyword: fat metabolism

Permethrin and ivermectin modulate in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 diabetes, which are closely associated with non-alcoholic fatty liver disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic . The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted fatty synthesis, while suppressed oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates fatty oxidation. Results in this study suggested that hepatic may be more susceptible to insecticide exposure in the presence of excessive fatty acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Effects of dietary perilla seed oil supplementation on , meat quality, and fatty profiles in Yellow-feathered chickens.

This study evaluated the effect of the dietary replacement of 1% lard (CT) with 1% perilla oil (PO), 0.9% perilla oil + 0.1% anise oil (PA), or 0.9% perilla oil + 0.1% ginger oil (PG) on indices of , antioxidant capacity, meat quality, and fatty profiles from Yellow-feathered chickens at day 63. Compared with the CT chickens, those given perilla oil had decreased (P < 0.05) plasma levels including triglycerides (TG), total cholesterol (TCH), and low-density lipoprotein cholesterol (LDL-C). Hepatic TG, TCH levels, and fatty synthase activity were also decreased (P < 0.05) in chickens fed diets containing perilla oil. Abdominal percentage was significantly decreased in birds fed the PG compared to CT diets. Birds fed the PA or PG diets had increased (P < 0.05) hepatic total SOD, glutathione peroxidase, and glutathione-S-transferase than in chickens given PO alone. In addition, the content of reduced glutathione (GSH) in breast muscle was lower (P < 0.05) in birds fed PO compared with those given PG, and the reverse was true for content of malondialdehyde. Compared with the CT diet, the PO diet decreased breast muscle shear values and increased yellowness (b*) of breast muscle (P < 0.05). Birds fed the PA or PG diets had meat with better overall acceptability than those fed the CT diet. Chickens fed perilla oil diets exhibited higher contents of α-linolenic (C18:3n-3), DHA (22:6n-3), polyunsaturated fatty acids, and n-3 fatty acids, together with a lower content of myristic (C14:0), (C16:0), stearic (C18:0), total saturated fatty acids, and n-6/n-3 ratio compared to controls (P < 0.05). These findings indicate that perilla oil has the potential to decrease -related indices and improve fatty profiles of breast meat in chickens without adverse effect on antioxidant status or meat quality; this was even better when perilla oil was given together with anise oil or ginger oil.© 2019 Poultry Science Association Inc.

Keyword: fat metabolism

Replication of Marek\'s Disease Virus Is Dependent on Synthesis of Fatty and Prostaglandin E.

Marek\'s disease virus (MDV) causes deadly lymphoma and induces an imbalance of the in infected chickens. Here, we discovered that MDV activates the fatty synthesis (FAS) pathway in primary chicken embryo fibroblasts (CEFs). In addition, MDV-infected cells contained high levels of fatty acids and showed increased numbers of droplets (LDs). Chemical inhibitors of the FAS pathway (TOFA and C75) reduced MDV titers by approximately 30-fold. Addition of the downstream metabolites, including malonyl-coenzyme A and , completely restored the inhibitory effects of the FAS inhibitors. Furthermore, we could demonstrate that MDV infection activates the COX-2/prostaglandin E (PGE) pathway, as evident by increased levels of arachidonic , COX-2 expression, and PGE synthesis. Inhibition of the COX-2/PGE pathway by chemical inhibitors or knockdown of COX2 using short hairpin RNA reduced MDV titers, suggesting that COX-2 promotes virus replication. Exogenous PGE completely restored the inhibition of the COX-2/PGE pathway in MDV replication. Unexpectedly, exogenous PGE also partially rescued the inhibitory effects of FAS inhibitors on MDV replication, suggesting that there is a link between these two pathways in MDV infection. Taken together, our data demonstrate that the FAS and COX-2/PGE pathways play an important role in the replication of this deadly pathogen. Disturbances of the in chickens infected with MDV contribute to the pathogenesis of disease. However, the role of in MDV replication remained unknown. Here, we demonstrate that MDV infection activates FAS and induces LD formation. Moreover, our results demonstrate that MDV replication is highly dependent on the FAS pathway and the downstream metabolites. Finally, our results reveal that MDV also activates the COX-2/PGE pathway, which supports MDV replication by activating PGE/EP2 and PGE/EP4 signaling pathways.Copyright © 2019 Boodhoo et al.

Keyword: fat metabolism

miR-21-5p regulates mitochondrial respiration and content in H9C2 cells.

Cardiovascular-related pathologies are the single leading cause of death in patients with chronic kidney disease (CKD). Previously, we found that a 5/6th nephrectomy model of CKD leads to an upregulation of miR-21-5p in the left ventricle, targeting peroxisome proliferator-activated receptor-α and altering the expression of numerous transcripts involved with fatty oxidation and glycolysis. In the present study, we evaluated the potential for knockdown or overexpression of miR-21-5p to regulate content, peroxidation, and mitochondrial respiration in H9C2 cells. Cells were transfected with anti-miR-21-5p (40 nM), pre-miR-21-5p (20 nM), or the appropriate scrambled oligonucleotide controls before treatment in culture or as part of the Agilent Seahorse XF fatty oxidation assay. Overexpression of miR-21-5p attenuated the -induced increase in cellular content, whereas suppression of miR-21-5p augmented it. The abundance of malondialdehyde, a product of peroxidation, was significantly increased with treatment in control cells but attenuated in pre-miR-21-5p-transfected cells. This suggests that miR-21-5p reduces oxidative stress. The cellular oxygen consumption rate (OCR) was increased in both pre-miR-21-5p- and anti-miR-21-5p-transfected cells. Levels of intracellular ATP were significantly higher in anti-mR-21-5p-transfected cells. Pre-miR-21-5p blocked additional increases in OCR in response to etomoxir and . Conversely, anti-miR-21-5p-transfected cells exhibited reduced OCR with both etomoxir and , and the glycolytic capacity was concomitantly reduced. Together, these results indicate that overexpression of miR-21-5p attenuates both content and peroxidation in H9C2 cells. This likely occurs by reducing cellular uptake and utilization, shifting cellular toward reliance on the glycolytic pathway. NEW & NOTEWORTHY Both overexpression and suppression of miR-21-5p augment basal and maximal mitochondrial respiration. Our data suggest that reliance on glycolytic and fatty oxidation pathways can be modulated by the abundance of miR-21-5p within the cell. miR-21-5p regulation of mitochondrial respiration can be modulated by extracellular lipids.

Keyword: fat metabolism

Novel hypolipidemic conjugates of fatty and bile with lysine for linkage.

Novel fatty -bile conjugates (1a-1k) were designed and synthesized by coupling of the fatty acids to the 3-OH of bile acids using lysine for linkage. In the conjugates, the 24-COOH of the bile acids was kept intact to preserve liver-specific recognition. The ability of the newly synthesized conjugates (at 100\u2009mg/kg dosage) to reduce total cholesterol (TC) and triglyceride (TG) levels in mice fed with high- diet (HFD) was evaluated. Conjugates of stearic with cholic and with ursodeoxycholic (at dosages of 50, 100, and 200\u2009mg/kg) were further evaluated to determine their ability to reduce aspartate aminotransferase (AST), alanine aminotransferase (ALT), TC, and TG levels in mice fed with HFD. All conjugates showed potent hypolipidemic activity. Further investigation revealed that compounds 1c and 1\u2009g not only dose-dependently reduced serum levels of TC and TG, but also inhibited the elevation of serum AST and ALT levels in mice fed with HFD. Thus, compounds 1c and 1\u2009g are promising hypolipidemic agents with hepatocyte protective effects against HFD-induced liver damage.

Keyword: fat metabolism

The mTORC1/4EBP1/PPARγ Axis Mediates Insulin-Induced Lipogenesis by Regulating Lipogenic Gene Expression in Bovine Mammary Epithelial Cells.

4EBP1 is a chief downstream factor of mTORC1, and PPARγ is a key lipogenesis-related transcription factor. mTORC1 and PPARγ are associated with . However, it is unknown which effector protein connects mTORC1 and PPARγ. This study investigated the interaction between 4EBP1 with PPARγ as part of the underlying mechanism by which insulin-induced synthesis and secretion are regulated by mTORC1 in primary bovine mammary epithelial cells (pBMECs). Rapamycin, a specific inhibitor of mTORC1, downregulated 4EBP1 phosphorylation and the expression of PPARγ and the following lipogenic genes: lipin 1, DGAT1, ACC, and FAS. Rapamycin also decreased the levels of intracellular triacylglycerol (TAG); 10 types of fatty ; and the accumulation of TAG, (PA), and stearic (SA) in the cell culture medium. Inactivation of mTORC1 by shRaptor or shRheb attenuated the synthesis and secretion of TAG and PA. In contrast, activation of mTORC1 by Rheb overexpression promoted 4EBP1 phosphorylation and PPARγ expression and upregulated the mRNA and protein levels of lipin 1, DGAT1, ACC, and FAS, whereas the levels of intracellular and extracellular TAG, PA, and SA also rose. Further, 4EBP1 interacted directly with PPARγ. Inactivation of mTORC1 by shRaptor prevented the nuclear location of PPARγ. These results demonstrate that mTORC1 regulates synthesis and secretion by inducing the expression of lipin 1, DGAT1, ACC, and FAS, which is likely mediated by the 4EBP1/PPARγ axis. This finding constitutes a novel mechanism by which synthesis and secretion are regulated in pBMECs.

Keyword: fat metabolism

Shengmai San Alleviates Diabetic Cardiomyopathy Through Improvement of Mitochondrial Metabolic Disorder.

Shengmai San (SMS), prepared from Panax ginseng, Ophiopogon japonicus, and Schisandra chinensisin, has been widely used to treat ischemic disease. In this study, we investigated whether SMS may exert a beneficial effect in diabetic cardiomyopathy through improvement of mitochondrial .A leptin receptor-deficient db/db mouse model was utilized, and lean age-matched C57BLKS mice served as non-diabetic controls. Glucose and profiles, myocardial structure, dimension, and function, and heart weight to tibial length ratio were determined. Myocardial ultrastructural morphology was observed with transmission electron microscopy. Protein expression and activity of oxidative phosphorylation (OXPHOS) complex were assessed using western blotting and microplate assay kits. We also observed cellular viability, mitochondrial membrane potential, OXPHOS complex activity, and cellular ATP level in -stimulated H9C2 cardiomyocytes. Changes in the sirtuin (SIRT1)/AMP-activated protein kinase (AMPK)/peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) pathway and mitochondrial uncoupling signaling were assessed using western blotting and quantitative real-time PCR.Leptin receptor-deficient db/db mice exhibit obesity, hyperglycemia, and hyperlipidemia, accompanied by distinct myocardial hypertrophy and diastolic dysfunction. SMS at a dose of 3 g/kg body weight contributed to a recovery of diabetes-induced myocardial hypertrophy and diastolic dysfunction. SMS administration led to an effective restoration of mitochondrial structure and function both in vivo and in vitro. Furthermore, SMS markedly enhanced SIRT1 and p-AMPKα protein levels and decreased the expression of acetylated-PGC-1α and uncoupling protein 2 protein. SMS also restored the depletion of NRF1 and TFAM levels in diabetic hearts and H9C2 cardiomyocytes.The results indicate that SMS may alleviate diabetes-induced myocardial hypertrophy and diastolic dysfunction by improving mitochondrial .© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fat metabolism

miR-125a-5p ameliorates hepatic glycolipid disorder in type 2 diabetes mellitus through targeting of STAT3.

Glycolipid metabolic disorder is an important cause for the development of type 2 diabetes mellitus (T2DM). Clarification of the molecular mechanism of metabolic disorder and exploration of drug targets are crucial for the treatment of T2DM. We examined miR-125a-5p levels in -induced AML12 cells and the livers of type 2 diabetic rats and mice, and then validated its target gene. Through gain- and loss-of-function studies, the effects of miR-125a-5p via targeting of STAT3 on regulating glycolipid were further illustrated and . We found that miR-125a-5p was significantly decreased in the livers of diabetic mice and rats, and STAT3 was identified as the target gene of miR-125a-5p. Overexpression of miR-125a-5p in C57BL/6 mice decreased STAT3 level and downregulated the expression levels of p-STAT3 and SOCS3. Consequently, SREBP-1c-mediated lipogenesis pathway was inhibited, and PI3K/AKT pathway was activated. Moreover, silencing of miR-125a-5p significantly increased the expression levels of STAT3, p-STAT3 and SOCS3, thus activating SREBP-1c pathway and suppressing PI3K/AKT pathway. Therefore, hyperglycemia, hyperlipidemia and decreased liver glycogen appeared in C57BL/6 mice. In -induced AML12 cells, miR-125a-5p mimic markedly increased glucose consumption and uptake and decreased the accumulation of droplets by regulating STAT3 signaling pathway. Consistently, miR-125a-5p overexpression obviously inhibited STAT3 expression in diabetic KK-Ay mice, thereby decreasing blood glucose and levels, increasing hepatic glycogen content, and decreasing accumulation of hepatic droplets in diabetic mice. Furthermore, inhibition of miR-125a-5p in KK-Ay mice aggravated glycolipid dysfunction through regulating STAT3. Our results confirmed that miR-125a-5p should be considered as a regulator of glycolipid in T2DM, which can inhibit hepatic lipogenesis and gluconeogenesis and elevate glycogen synthesis by targeting STAT3.

Keyword: fat metabolism

Dual Specificity Phosphatase 12 Regulates Hepatic Through Inhibition of the Lipogenesis and Apoptosis Signal-Regulating Kinase 1 Pathways.

Nonalcoholic fatty liver disease (NAFLD) has become the most common cause of chronic liver disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and inflammation in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high- diet (HFD)-induced and high- high-cholesterol diet-induced hyperinsulinemia and liver steatosis and decreased insulin sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, insulin resistance, and inflammation. At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic . DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit lipogenesis under high- conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.

Keyword: fat metabolism

Alterations to the microbiota-colon-brain axis in high--diet-induced obese mice compared to diet-resistant mice.

Obesity is underpinned by both genetic and environmental factors, including a high-saturated- diet. Some mice develop diet-induced obesity (DIO), but others remain diet resistant (DR) despite intake of the same high-saturated- diet, a phenomenon that mimics characteristics of the human obese phenotype. Microbiota-colon-brain axis regulation is important for energy and cognition. Using DIO and DR mouse models, this study aimed to examine gut microbiota, colonic inflammation and cognitive function to elucidate the role of microbiota-gut-brain regulation in DIO. C57Bl6/J mice fed a chronic saturated- diet for 22 weeks showed significant body weight gain differences, with the top one third gaining 48% heavier body weight than the lower one third. There was significant reduction in gut microbiota richness and diversity in DIO mice but not in DR mice. At the phylum level, DIO mice had increased abundance of Firmicutes and Antinobacteria, and decreased abundance of Bacterioides and Proteobacteria in gut microbiota. DIO mice exhibited reduced tight junction proteins, increased plasma endotoxin lipopolysaccharide (LPS) and increased inflammation in the colon and liver. Recognition memory and spatial memory were impaired in DIO mice, associated with decreased Bacteroidetes. Further examination showed that hippocampal brain-derived neurotrophic factor was significantly decreased in DIO mice (vs. DR). Conversely, DR mice showed no changes in the above parameters measured. Therefore, gut microbiota, colon inflammation and circulating LPS may play a major role in the development of the obese phenotype and cognitive decline associated with a chronic high-saturated- diet.Copyright © 2018. Published by Elsevier Inc.

Keyword: fat metabolism

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic fatty liver disease (NAFLD), and type 2 diabetes (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 species were identified, quantified, and classified into free fatty acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of fatty acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

Keyword: fat metabolism

Cyclophilin D participates in the inhibitory effect of high- diet on the expression of steroidogenic acute regulatory protein.

The high- diet (HFD)-induced obesity is responsible for the testosterone deficiency (TD). However, the mechanism remains unknown. Mitochondrial homeostasis is proved to be important for maintaining the function of steroidogenic acute regulatory protein (StAR), the first rate-limiting enzyme in testosterone synthesis. As the key regulator of mitochondrial membrane permeability, cyclophilin D (CypD) plays a crucial role in maintaining mitochondrial function. In this study, we sought to elucidate the role of CypD in the expression of StAR affected by HFD.To analyse the influence of CypD on StAR in vivo and in vitro, mouse models of HFD, CypD overexpression and CypD knockout (Ppif ) as well as Leydig cells treated with (PA) and CypD overexpression plasmids were examined with an array of metabolic, mitochondrial function and molecular assays.Compared with the normal diet mice, consistent with reduced testosterone in testes, the expressions of StAR in both mRNA and protein levels in HFD mice were down-regulated, while expressions of CypD were up-regulated. High- intake impaired mitochondrial function with the decrease in StAR in Leydig cells. Overexpression of CypD inhibited StAR expressions in vivo and in vitro. Compared with C57BL/6 mice with HFD, expressions of StAR were improved in Ppif mice with HFD.Mitochondrial CypD involved in the inhibitory effect of HFD on StAR expression in testes.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fat metabolism

Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

The fetus is dependent on delivery of fatty acids (FAs) by the syncytiotrophoblast, the transporting epithelium of the human placenta. Obese pregnant women have dyslipidemia; however, whether obesity impacts placental transport and remains to be fully established. Palmitoleic (POA), an FA with anti-inflammatory and insulin-sensitizing properties, is synthesized from (PA) catalyzed by stearoyl-coenzyme A desaturase (SCD) activity. We hypothesized that the uptake and incorporation of FAs and POA synthesis are reduced in primary human trophoblasts (PHTs) isolated from pregnancies complicated by maternal obesity. Villous cytotrophoblasts were isolated from 7 placentas of obese [body mass index (BMI) = 37.5 ± 1.9] and 12 normal (BMI = 23.6 ± 0.6) mothers. FA uptake and incorporation were assessed using uniformly labeled (U[C])-FA mixtures of PA, oleic (OA), linoleic , and docosahexaenoic . Cellular [C] FAs were quantified both in total cellular lipids and in classes by GC-MS. Uptake and incorporation of [C] FAs in total cellular lipids were not different in PHTs isolated from obese mothers compared with normal mothers. Only the concentration of OA was increased in the triglyceride fraction ( < 0.05) if the mother was obese. We found an isotopic enrichment of POA after U[C]-PA treatment, demonstrating SCD activity in PHT cells. Labeled POA content and the POA:PA ratio were significantly lower in PHTs isolated from placentas of obese mothers compared with normal, healthy controls. Decreased syncytiotrophoblast POA synthesis may contribute to insulin resistance and low-grade inflammation in the mother, placenta, or fetus (or a combination of the 3) in pregnancies complicated by obesity.-Ferchaud-Roucher, V., Barner, K., Jansson, T., Powell, T. L. Maternal obesity results in decreased syncytiotrophoblast synthesis of palmitoleic , a fatty with anti-inflammatory and insulin-sensitizing properties.

Keyword: fat metabolism

Deciphering the intervention mechanism of Taohong Siwu Decoction following the abnormal uterine bleeding rats based on serum metabolic profiles.

Abnormal uterine bleeding (AUB), one of the most significant characters of incomplete abortion, is a widespread phenomenon in gynecological that put a woman into a terrible physiological and psychological state. Taohong Siwu Decoction (TSD) is a traditional Chinese medicine (TCM) prescriptions which have treated AUB in China for decades. Our previous study elucidated that TSD reduced the volume of uterine bleedings as well as repaired the endometrium. The present study aims to investigate the mechanisms of TSD on AUB based on serum metabolomics. In this study, serum metabolic profile data was collected using ultra high-performance liquid chromatography with ion trap/time-of-flight mass spectrometry and gas chromatography-mass spectrometry. 23 potential biomarkers (urea, serine, L-proline, L-glutamic , , l-acetylcarnitine, LysoPC(16:0), LysoPC(20:4), l-proline, linoleic , stearic , l-isoleucine, phenylalanine, l-tyrosine, Oleic , et al) were eventually identified using multivariate statistical analysis (PCA and OPLS-DA) with VIP\u2009>\u20091, P\u2009<\u20090.05. Correlation analysis, fold-change (FC), area under receiver characteristic (ROC), false discovery rate (FDR) were used for data confirmation to ensure the authenticity of the data. The related-metabolic pathway mainly included amino (Phenylalanine, tyrosine, and tryptophan ; Valine, leucine and isoleucine biosynthesis; Arginine and proline ; Glycine, serine and threonine ) and (linoleic , glycerophospholipid ). The results show that TSD has a favorable therapeutic effect on AUB by adjusting the metabolic disorders, which could provide dietary guidance for the clinic.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Protective effect and mechanism of Qiwei Tiexie capsule on 3T3-L1 adipocytes cells and rats with nonalcoholic fatty liver disease by regulating LXRα, PPARγ, and NF-κB-iNOS-NO signaling pathways.

Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic liver disease and nonalcoholic fatty liver disease (NAFLD).This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD.The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and oxidation were assessed by 2-deoxy-D-[H] glucose and [1-C] , respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), liver X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of emulsion and sucrose for 9 weeks. The effects of QWTX on , liver function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee fatty (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the liver, as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected.QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the CO production rate to 4.15, which indicated the reducing the fatty accumulation. In NAFLD, QWTX attenuated liver steatosis, vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In liver tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model.QWTX protected the liver injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways.Copyright © 2019. Published by Elsevier B.V.

Keyword: fat metabolism

APOE and the Association of Fatty Acids With the Risk of Stroke, Coronary Heart Disease, and Mortality.

Background and Purpose- The role of dietary on cardiovascular health and mortality remains under debate. Because the APOE is central to the transport and of lipids, we examined associations between plasma fatty acids and the risk of stroke, coronary heart disease, and mortality by APOE-ε4 genotype. Methods- We included 943 FHS (Framingham Heart Study) and 1406 3C (Three-City) Bordeaux Study participants. Plasma docosahexaenoic, linoleic, arachidonic, and fatty acids were measured at baseline by gas chromatography. All-cause stroke, ischemic stroke, coronary heart disease, and all-cause mortality events were identified prospectively using standardized protocols. Each cohort used Cox models to separately relate fatty levels to the risk of developing each event during ≤10 years of follow-up adjusting for potential confounders and stratifying by APOE genotype (ε4 carriers versus noncarriers). We then meta-analyzed summary statistics using random-effects models. Results- On average, participants had a mean age of 74 years, 61% were women, and 21% (n=483) were APOE-ε4 carriers. Meta-analysis results showed that, only among APOE-ε4 carriers, every SD unit increase in linoleic was associated with a reduced risk of all-cause stroke (hazard ratio [HR], 0.54 [95% CI, 0.38-0.78]), ischemic stroke (HR, 0.48 [95% CI, 0.33-0.71]), and all-cause mortality (HR, 0.70 [95% CI, 0.57-0.85]). In contrast, every SD unit increase in was related to an increased risk of all-cause stroke (HR, 1.58 [95% CI, 1.16-2.17]), ischemic stroke (HR, 1.76 [95% CI, 1.26-2.45]), and coronary heart disease (HR, 1.48 [95% CI, 1.09-2.01]), also in APOE-ε4 carriers only. Results for docosahexaenoic and arachidonic were heterogeneous between cohorts. Conclusions- These exploratory results suggest that APOE-ε4 carriers may be more susceptible to the beneficial or adverse impact of fatty acids on cardiovascular disease and mortality. In this subgroup, higher linoleic was protective for stroke and mortality, whereas was a risk factor for stroke and coronary heart disease. The mechanisms underlying these novel findings warrant further investigation.

Keyword: fat metabolism

Exploring the interactions between serum free fatty acids and fecal microbiota in obesity through a machine learning algorithm.

Serum free fatty acids (FFA) are generally elevated in obesity. The gut microbiota is involved in the host energy through the regulation of body storage, and a link between diet, FFA and the intestinal microbiota seems to exist. Our aim was to explore the interaction among serum FFA levels, gut microbiota, diet and obesity through a model regression tree in 66 subjects (age 52.7\u202f±\u202f11.2\u202fy) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. Microbiota was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. , Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA\u202f>\u202f0.235\u202fμg/mL and Bacteroides\u202f>\u202f9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

[Psoralen and isopsoralen improve disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using ( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Keyword: fat metabolism

Lipotoxicity reduces β cell survival through islet stellate cell activation regulated by -related molecules.

Islet stellate cells (ISCs) activation is mainly associated with islet fibrosis, which contributes to the progression of type 2 diabetes. However, the molecular mechanism underlying this process is not fully understood.In order to investigate this process the current study examined ectopic accumulation in rats with high- diet (HFD) induced obesity. Levels of lipotoxicity-induced ISC activation and islet function were assessed via intraperitoneal glucose and insulin tolerance tests, and immunohistochemistry. The expression of - and ISC activation-related markers was evaluated in cultured ISCs treated with (PA) using quantitative PCR and western blotting. We also overexpressed sterol regulatory element-binding protein (SREBP)-1c in ISCs by lentiviral transduction, and assessed the effects on insulin release in co-cultures with isolated rat islets.HFD increased body weight and ectopic accumulation in pancreatic islets. Lipotoxicity caused progressive glucose intolerance and insulin resistance, upregulated α-smooth muscle actin, and stimulated the secretion of extracellular matrix. Lipotoxicity reduced the expression of -related molecules in ISCs treated with PA, especially SREBP-1c. Overexpression of SREBP-1c in ISCs improved islet viability and insulin secretion in co-cultures.These results indicate that lipotoxicity-induced ISC activation alters islet function via regulation of , suggesting that therapeutic strategies targeting activated ISC may be an effective treatment for prevention of ISC activation-associated islet dysfunction.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Physicochemical characteristics of blend from hydrogenated coconut oil and acyl migrated palm mid-fraction.

Palm mid-fraction (PMF), which has a high content of symmetric POP, was converted to asymmetric PPO (APMF) via acyl migration. After solvent fractionation, the liquid phase of acyl migrated PMF (APMF-L) was obtained and blended with hydrogenated coconut oil (HCO, 50:50, w/w) to produce a blend (namely, an alternative blend) which had reduced saturated fatty content while having similar melting behavior to HCO. In an alternative blend, the major fatty acids were lauric (27.94), (26.93) and oleic (15.75\u202fmol%) . The solid index was quite similar to that of HCO, especially at 28-44\u202f°C. Nevertheless, an alternative blend had lower saturated fatty content, by 18%, compared to HCO. The content of highly atherogenic myristic was reduced by approximately 40%. The alternative blend in this study could be used as a raw material for non-dairy cream with low saturated content.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Analysis of Titin in Red and White Muscles: Crucial Role on Muscle Contractions Using a Fish Model.

Several studies have compared molecular components between red and white skeletal muscles in mammals. However, mammalian skeletal muscles are composed of mixed types of muscle fibers. In the current study, we analyzed and compared the distributions of titin, , phosphate ions, and fatty levels in red and white muscles using a fish model (), which is rich in red and white muscles, and these are well separated. Oil-red O staining showed that red muscle had more-abundant lipids than did white muscle. A time-of-flight secondary-ion mass spectrometric (TOF-SIMS) analysis revealed that red muscle possessed high levels of and oleic , but white muscle contained more phosphate ions. Moreover, elastica-van Gieson (EVG) and Mito-Tracker green FM staining showed that collagen and elastic fibers were highly, respectively, distributed in connective tissues and mitochondria in red muscle. An electron micrographic analysis indicated that red muscle had a relatively higher number of mitochondria and longer sarcomere lengths and Z-line widths, while myofibril diameters were thicker in white muscle. Myofibrillar proteins separated by SDS-PAGE showed that the major giant protein, titin, was highly expressed in white muscle than in red muscle. Furthermore, ratios of titin to myosin heavy chain (MHC) (titin/MHC) were about 1.3 times higher in white muscle than red muscle. We postulated that white muscle is fit for short and strong contractile performance due to high levels of titin and condensed sarcomeres, whereas red muscle is fit for low intensity and long-lasting activity due to high levels of lipids and mitochondria and long sarcomeres.

Keyword: fat metabolism

Thioacetamide potentiates high cholesterol and high diet induced steato-hepatitic changes in livers of C57BL/6J mice: A novel eight weeks model of fibrosing NASH.

There is an inadequacy of relevant animal models to study non-alcoholic steatohepatitis (NASH) and fibrosis. Here, we co-administered thioacetamide (TH) along with fast food diet (FFD) to C57BL/6\u2009J mice for eight weeks. The treatments were: a) standard chow, SC b) FFD c) FFD\u2009+\u2009TH [75\u2009mg/kg], FTH d) SC\u2009+\u2009TH [150\u2009mg/kg], STH for 8 weeks. In in-vitro model, Hep3B cells were exposed to (PA) and TH viz. PA (0.25\u2009mM) + TH (25\u2009mM), PA (0.5\u2009mM) alone and TH (50\u2009mM) alone for 12\u2009h, later supernatant media was transferred to LX-2 cells, for another 12\u2009h. Molecular and cellular events related to inflammation, fibrosis, collagen deposition were studied. The FTH mice featured hepatic inflammation, severe diffuse fibrosis, and collagen deposition, which were less severe in FF & STH groups. In FTH group the protein expressions of α-SMA, TGF-ß, Col1\u2009A1, CYP2E1, were up-regulated as compared to the FF group. The in-vivo findings were complemented in the LX-2 and Hep3B cells. The protein expressions of inflammatory and cellular injury markers were significantly higher in PA\u2009+\u2009TH exposed LX-2 cells. This novel model manifested hepatic inflammation and fibrosis in just eight weeks, which may be exploited for rapid screening of novel anti-NAFLD and liver anti-fibrotic agents.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

A high- diet induces rapid changes in the mouse hypothalamic proteome.

Prolonged over-consumption of a high- diet (HFD) commonly leads to obesity and insulin resistance. However, even 3\u2009days of HFD consumption has been linked to inflammation within the key homeostatic brain region, the hypothalamus.Mice were fed either a low- diet (LFD) or HFD containing 10% or 60% (Kcal) respectively from for 3\u2009days. Mice were weighed, food intake measured and glucose tolerance calculated using intraperitoneal glucose tolerance tests (IPGTT). Proteomic analysis was carried out to determine if hypothalamic proteins were changed by a HFD. The direct effects of dietary fatty acids on mitochondrial morphology and on one of the proteins most changed by a HFD, dihydropyrimidinase-related protein 2 (DRP-2) a microtubule-associated protein which regulates microtubule dynamics, were also tested in mHypoE-N42 (N42) neuronal cells challenged with (PA) and oleic (OA).Mice on the HFD, as expected, showed increased adiposity and glucose intolerance. Hypothalamic proteomic analysis revealed changes in 104 spots after 3\u2009days on HFD, which, when identified by LC/MS/MS, were found to represent 78 proteins mainly associated with cytoskeleton and synaptic plasticity, stress response, glucose and mitochondrial function. Over half of the changed proteins have also been reported to be changed in neurodegenerative conditions such as Alzheimer\'s disease. Also,in N42 neurons mitochondrial morphology and DRP-2 levels were altered by PA but not by OA.These results demonstrate that within 3\u2009days, there is a relatively large effect of HFD on the hypothalamic proteome indicative of cellular stress, altered synaptic plasticity and mitochondrial function, but not inflammation. Changes in N42 cells show an effect of PA but not OA on DRP-2 and on mitochondrial morphology indicating that long-chain saturated fatty acids damage neuronal function.

Keyword: fat metabolism

Prepartum fatty supplementation in sheep. IV. Effect of calcium salts with eicosapentaenoic and docosahexaenoic in the maternal and finishing diet on lamb liver and adipose tissue during the lamb finishing period1.

The objective of this study was to evaluate the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) supplementation to ewes during late gestation on finishing lamb liver and adipose tissue fatty (FA) profile and gene expression. Lambs born from ewes supplemented with Ca salts of EPA + DHA, or palm FA distillate (PFAD) high in and oleic at 0.39% DM during the last 50 d of gestation were used. Lambs were weaned at 61 d of age and adapted to a high concentrate diet for 1.5 mo. After adaptation, 74 lambs (28 pens) were blocked by sex and BW and used in a 2 × 2 factorial arrangement of treatments using the factors of dam supplementation (DS) and lamb supplementation (LS) of Ca salts of EPA + DHA or PFAD at 1.48% DM. Lambs were slaughtered after 42 d and liver and adipose tissue collected for FA and gene expression analysis. Liver concentrations of EPA and DHA were greater (P < 0.01) with LS of EPA + DHA vs. PFAD during the finishing period. In adipose tissue, a lamb × dam interaction was observed for EPA (P = 0.02) and DHA (P = 0.04); LS of EPA + DHA increased EPA and DHA, but the increase was greatest in lambs born from ewes supplemented with PFAD. No lamb × dam treatment interactions were observed for gene expression in liver tissue (P > 0.10). Hepatic mRNA abundance of hormone-sensitive lipase (HSL; P = 0.01) was greater in lambs born from EPA + DHA ewes vs. lambs from PFAD ewes. mRNA expression of stearoyl-CoA desaturase (P < 0.01), fatty synthase (P = 0.01), Δ5-desaturase (P < 0.01), and Δ6-desaturase (P < 0.01) were decreased in liver of EPA + DHA lambs. A significant lamb × dam diet interaction was observed for elongation of very long chain fatty 2 in adipose tissue (P = 0.01); lambs supplemented with the same FA as their dams had lower expression. Expression of HSL tended (P = 0.08) to be decreased in adipose of EPA + DHA lambs born from EPA + DHA ewes. The changes in mRNA expression suggest that lipogenesis decreased, and lipolysis increased in lamb liver with EPA + DHA vs. PFAD supplementation during the finishing period. In adipose tissue, changes suggest that lipogenesis decreased in lambs born from EPA + DHA supplemented dams and supplemented with EPA + DHA during the finishing period. In addition, these results suggest an interaction between supplementation of FA to dams during late gestation on lamb response of adipose tissue, but not liver, to FA supplementation during the finishing period.© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: fat metabolism

Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by .

Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased , protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug and antioxidant adaptation in HepG2 cells treated with , which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and .

Keyword: fat metabolism

Adverse Effects in Skeletal Muscle Following the Medicinal Use of Nicotiana glauca.

Nicotiana glauca is a cosmopolitan shrub, used in medicine to treat swellings, wounds, sores and cancer. However, its users lack of knowledge of the adverse effects. We seek to evaluate the effects of extracts from N. glauca on myoblasts, identifying the compounds which cause undesirable effects. Myoblasts are important in muscle homeostasis, thus a high death rate of them cause myopathies. We performed an ethanolic extraction from leaves of N. glauca and the extract was successively partitioned with hexane, chloroform and ethyl acetate. The effects of extracts in C2C12 cells were analysed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), Mitotracker and 4\',6-diamidino-2-phenylindole (DAPI) staining, Western blotting, real-time PCR and immunofluorescence assays. Caspase activity was studied. The fraction with the highest apoptotic effects was analysed by chromatography, NMR and GC-MS spectrometry were used to identify the apoptotic agent, after which its biological activity was evaluated. The extracts from N. glauca induced apoptosis in C2C12 cells involving caspase-3/7. We found that the extracts trigger a defence response in muscle through Akt and heat shock protein 27 (HSP27). We identified an apoptotic agent as . These data suggest that the use of N. glauca in hormone replacement therapy, or in other therapies affects skeletal muscle homeostasis, worsening the negative effects of the menopause. Thus, the relevance of this work lies in the fact that it is the first time that a report about the molecular mechanism responsible for the side effects of medicinal use of N. glauca, has been shown. Moreover the compound responsible for these effects has been identified.

Keyword: fat metabolism

Human cytochrome P450 enzymes bind drugs and other substrates mainly through conformational-selection modes.

Cytochrome P450 (P450) enzymes are major catalysts involved in the oxidations of most drugs, steroids, carcinogens, -soluble vitamins, and natural products. The binding of substrates to some of the 57 human P450s and other mammalian P450s is more complex than a two-state system and has been proposed to involve mechanisms such as multiple ligand occupancy, induced-fit, and conformational-selection. Here, we used kinetic analysis of binding with multiple concentrations of substrates and computational modeling of these data to discern possible binding modes of several human P450s. We observed that P450 2D6 binds its ligand rolapitant in a mechanism involving conformational-selection. P450 4A11 bound the substrate lauric via conformational-selection, as did P450 2C8 with . Binding of the steroid progesterone to P450 21A2 was also best described by a conformational-selection model. Hexyl isonicotinate binding to P450 2E1 could be described by either a conformational-selection or an induced-fit model. Simulation of the binding of the ligands midazolam, bromocriptine, testosterone, and ketoconazole to P450 3A4 was consistent with an induced-fit or a conformational-selection model, but the concentration dependence of binding rates for varying both P450 3A4 and midazolam concentrations revealed discordance in the parameters, indicative of conformational-selection. Binding of the P450s 2C8, 2D6, 3A4, 4A11, and 21A2 was best described by conformational-selection, and P450 2E1 appeared to fit either mode. These findings highlight the complexity of human P450-substrate interactions and that conformational-selection is a dominant feature of many of these interactions.© 2019 Guengerich et al.

Keyword: fat metabolism

Mechanisms of electrical remodeling in lipotoxic guinea pig heart.

To develop an adult guinea pig model of lipotoxicity and explore the underlying mechanisms associated with changes in the expression of the delayed rectifier potassium current (I).Lipotoxicity may represent a common link among metabolic disorders and a higher vulnerability to arrhythmias.Whole-cell patch clamp, and (PA, a potent inducer of lipotoxicity), were used to assess mechanisms of short-term (∼50 days) high- diet (HFD) feeding on atrial electrophysiology in guinea pig hearts and myocytes.HFD fed guinea pigs were significantly heavier, displayed hypertriglyceridemia and hypercholesterolemia; but no signs of hyperglycemia or inflammation compared to low- diet fed controls. Increasing cardiac PA levels, resulted in shortened atrial action potential duration, and increased I density. Inhibition of phosphoinositide 3-kinase (PI3K) prevented increases in I due to PA. Acute (≥1hr) exposure of atrial myocytes to exogenous PA (1\u202fmM) increased the density of the rapid delayed rectifier potassium current I, while it was decreased with the unsaturated oleic (OA, 1\u202fmM). Serine-threonine protein phosphatase-2 (PP2A) inhibition with cantharidin reversed the effect of OA on I.Our data provide evidence of a novel lipotoxic guinea pig model with signs of vulnerability to arrhythmias. Inhibition of PA/PI3K/I and/or activation of the OA/PP2A/I pathways may be therapeutically beneficial for lipotoxic arrhythmias.Copyright © 2019 The Author. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

The inhibition of Nrf2 accelerates renal deposition through suppressing the ACSL1 expression in obesity-related nephropathy.

Obesity has become a worldwide epidemic, and the incidence of obesity is increasing year by year. Obesity-related nephropathy (ORN) is a common kidney complication of obesity. Long-chain acyl-CoA synthetases-1, (ACSL1), is a key enzyme in the oxidative of fatty acids in mitochondria and ACSL1 may play a direct role in renal deposition and promote the progress of ORN. In this study, we focus on the renoprotective role of ACSL1 in ORN. Electron microscopy, immunohistochemical (IHC) staining, Western blot, and real-time PCR were used to detect the expression of ACSL1and Nrf2 in ORN patients, ob/ob mice and (PA)-treated HK-2 cells. Oil red staining and Elisa Kit were used to detect the intracellular FFA and TG contents in ob/ob mice and PA-treated HK-2 cells. Dihydroethidium (DHE) staining and the MDA/SOD measurement were used to detect the ROS production. In order to demonstrate the role of ACSL1 and the interaction between ACSL1 and Nrf2 in ORN, related siRNA and plasmid were transfected into HK-2 cells. More ROS production and renal deposition have been found in ORN patients, ob/ob mice and PA-treated HK-2 cells. Compared with control, all the expression of ACSL1and Nrf2 were down-regulated in ORN patients, ob/ob mice and PA-treated HK-2 cells. The Nrf2 could regulate the expression of ACSL1 and the ACSL1 played the direct role in renal deposition. The Nrf2 is inhibited in ORN, resulting more ROS production and oxidative stress. Increased oxidative stress will suppress the expression of ACSL1, which could increase the intracellular FFA and TG contents, ultimately leading to renal deposition in renal tubulars and accelerating the development of ORN.

Keyword: fat metabolism

Inhibition of soluble epoxide hydrolase attenuates a high- diet-mediated renal injury by activating PAX2 and AMPK.

A high- diet (HFD) causes obesity-associated morbidities involved in macroautophagy and chaperone-mediated autophagy (CMA). AMPK, the mediator of macroautophage, has been reported to be inactivated in HFD-caused renal injury. However, PAX2, the mediator for CMA, has not been reported in HFD-caused renal injury. Here we report that HFD-caused renal injury involved the inactivation of Pax2 and Ampk, and the activation of soluble epoxide hydrolase (sEH), in a murine model. Specifically, mice fed on an HFD for 2, 4, and 8 wk showed time-dependent renal injury, the significant decrease in renal Pax2 and Ampk at both mRNA and protein levels, and a significant increase in renal sEH at mRNA, protein, and molecular levels. Also, administration of an sEH inhibitor, 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl)urea, significantly attenuated the HFD-caused renal injury, decreased renal sEH consistently at mRNA and protein levels, modified the renal levels of sEH-mediated epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs) as expected, and increased renal Pax2 and Ampk at mRNA and/or protein levels. Furthermore, (PA) treatment caused significant increase in , and decrease in both Pax2 and Ampk in murine renal mesangial cells (mRMCs) time- and dose-dependently. Also, 14(15)-EET (a major substrate of sEH), but not its sEH-mediated metabolite 14,15-DHET, significantly reversed PA-induced increase in , and PA-induced decrease in Pax2 and Ampk. In addition, plasmid construction revealed that Pax2 may positively regulate Ampk transcriptionally in mRMCs. This study provides insights into and therapeutic target for the HFD-mediated renal injury.

Keyword: fat metabolism

[Laurine fatty acids, medium fatty acids and triglycerides, hyperlipidemia, resistance to insulin, prevention of atherosclerosis and ateromatosis.]

Although the biochemistry of the positive effects of medium-chain fatty acids (FA) and triglycerides (TG) of the same name in vivo is not fully understood, food enriched with medium-chain LC and the same TG is effective in patients with type I diabetes, insulin resistance syndrome and in neurodegenerative pathology. Lauric C12 LC is half the FA in coconut oil. Residents of southeast Asia with constant use of coconut oil, have a low level of diseases of the cardiovascular system in the population. With a regulatory intake with food C12:0 laurin FA formed moderate ketosis and neuroprotective effect. Unlike long-chain LC, medium-chain TG cells are not deposited either in visceral cells, or in insulin-dependent adipocytes. Medium-chain fatty acids rapidly oxidize mitochondria; the formation of acetyl-CoA cells is used to form ketone bodies, activating thermogenesis in orange and brown adipocytes. Experiments with animals and observations in the clinic showed that taking medium-chain TG with food is more physiological than long-chain oils. This significantly increases the level of cholesterol in high-density lipoproteins. Food enriched with medium chain TG is optimal for increasing the ketone content in blood plasma, cerebrospinal fluid without limiting the carbohydrate content in food. The formation of excess ketone bodies by cells can be achieved by activating the metabolic transformations of medium-chain FAs, without fasting and preserving carbohydrates in food. Coconut oil has a positive effect on the cardiovascular system, preventing the formation of atherosclerosis and atheromatosis. Effective in the prevention of the pathology of the cardiovascular system is a decrease in food amounts of , an increase in oleic , polyene FA with a simultaneous increase in the proportion of medium-chain FA.

Keyword: fat metabolism

Outdoor cultivation of the green microalga Chlorella vulgaris under stress conditions as a feedstock for biofuel.

The present work investigated the potential of the green alga Chlorella vulgaris to produce high-quality biofuel under culture stress conditions. The cultivation was carried out in a 1000 l\xa0open plate tank system, which provides biomass yields comparable to open pond systems, but with less area needed. Algal biomass and content were measured repeatedly. We compared the two solvent systems n-hexane and hexane/isopropanol (HIP) for extraction efficiency of lipids and applied three different extraction methods Soxhlet, soaking, and soaking followed by Soxhlet (soak-Sox). The combination of the HIP solvent and the soak-Sox provided the highest yield (15.8\u2009±\u20090.174). Volumetric biomass and productivity were 0.201\xa0g\xa0l\xa0day and 31.71\xa0mg\xa0l\xa0day, respectively, whereas areal biomass and productivity were 25.73\xa0g\xa0m\xa0day and 4.066\xa0g\xa0m\xa0day, respectively. The fatty profile by means of gas chromatography resulted in seven fatty acids from C to C. The most abundant fatty methyl esters (FAMES) were (C16:0), oleic (C18:1), and stearic (C18:0) acids. synthesis enhanced by optimizing the Kuhl growth medium with replacing nitrate by urea (50% N compared to the original recipe) increased salt content (10\xa0g/l NaCl), ferrous sulfate (0.5\xa0g/l), and sodium acetate addition (1\xa0g/l). With regard to density, kinematic viscosity, gravity, pour point, flash point, and cetane number, the Chlorella-biodiesel comply with ASTM and EN standards thus pointing at the high potential of lipids synthesized by Chlorella as a feedstock for biodiesel production.

Keyword: fat metabolism

[Exploration about the protection mechanism of 5-hydroxy-1-methylhydantoin on paraquat poisoning model].

To investigate the effects of 5-hydroxy-1-methylhydantoin (HMH) on kidney injury induced by paraquat (PQ).Fifteen SPF healthy Kunming mice were randomly divided into normal saline (NS) control group, PQ poisoning model group and HMH intervention group, with 5 mice in each group. PQ poisoning model was challenged by one-time gavage of 30 mg/kg PQ solution. The NS group received the same amount of NS by gavage. The HMH group was given 100 mg/kg of HMH immediately after the model was made and continued to be gavaged. Mice in each group were sacrificed 1 day after HMH gavage and heart blood and renal tissue were harvested for examination. The morphological changes of renal tissue were observed under light microscope by hematoxylin-eosin (HE) staining. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in renal tissue were detected according to the instructions of the kit. The expression of heme oxygenase-1 (HO-1) and interleukin-1β (IL-1β) in renal tissues were detected by Western Blot. The serum metabolites were detected by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS), the overall distribution of each sample was observed by principal component analysis (PCA), the accuracy of the model was evaluated by multidimensional analysis orthogonal partial least squares-discriminant analysis (OPLS-DA), and the difference metabolites were screened by variable importance in the projection (VIP) value > 1.Light microscopic observation showed that: glomerular structure in NS group was clear, there was no hyperemia and inflammatory cell infiltration in renal interstitium and blood vessels. In PQ group, some glomeruli atrophy and necrosis, capillary congestion in glomeruli, infiltration of inflammatory cells around glomeruli, swelling of renal tubular epithelial cells, slight stenosis of lumen, and occasional necrosis and exfoliation of epithelial cells occurred. The degree of kidney injury in HMH group was significantly less than that in PQ group. Compared with the NS group, the content of MDA in the PQ group was significantly increased (nmol/g: 6.70±0.84 vs. 2.70±0.43, P < 0.01) and the activity of SOD was significantly decreased (kU/L: 33.30±4.66 vs. 50.20±3.23, P < 0.05), the protein expression of HO-1 and IL-1β were significantly increased (HO-1/β-actin: 1.11±0.12 vs. 0.61±0.13, IL-1β/β-actin: 0.93±0.13 vs. 0.32±0.06, both P < 0.05). Compared with the PQ group, the content of MDA in the HMH group was significantly decreased (nmol/g: 5.10±0.93 vs. 6.70±0.84, P < 0.05) and the activity of SOD was significantly increased (kU/L: 61.00±9.02 vs. 33.30±4.66, P < 0.05), the protein expression of HO-1 was significantly decreased (HO-1/β-actin: 0.77±0.07 vs. 1.11±0.12, P < 0.05), however, there was no significant difference in the protein expression of IL-1β (IL-1β/β-actin: 0.87±0.13 vs. 0.93±0.13, P > 0.05). Metabolite detection results showed that: compared with NS group, the levels of creatinine, glycine, succinic , fumaric and citric were significantly increased in the PQ group (VIP value was 1.50, 1.58, 1.64, 1.74 and 1.95 respectively, all P < 0.05), while the levels of ,α-tocopherol and 6-phosphogluconic were significantly decreased (VIP value was 1.10, 1.55 and 1.56 respectively, all P < 0.05). Compared with the PQ group, the levels of creatinine and citric were significantly decreased in the HMH group (VIP value was 1.50 and 1.86, both P < 0.05), while trans-4-hydroxy-proline, D-glyceric , 2, 6-fructose phosphate, 6-phosphate gluconic and aminomalonic were significantly increased (VIP value was 1.36, 1.55, 1.63, 1.68 and 1.76 respectively, all P < 0.05).HMH protects kidney injury caused by PQ poisoning by correcting tricarboxylic acids cycle disturbance, peroxidation and energy disturbance, and its mechanism is related to the regulation of HO-1 protein expression through Nrf2 pathway.

Keyword: fat metabolism

Berberine attenuates nonalcoholic hepatic steatosis through the AMPK-SREBP-1c-SCD1 pathway.

Berberine (BBR), a natural compound extracted from Chinese herb, has been shown to effectively attenuate nonalcoholic fatty liver disease (NAFLD) in clinic. However, the mechanism underlying the effect of BBR is not fully understood. Stearyl-coenzyme A desaturase 1 (SCD1) mediates in liver. Therefore, we hypothesized that SCD1 mediated the beneficial effect of BBR on NAFLD.The expression of SCD1 was measured in the liver of NAFLD patients and ob/ob mice. The effect of BBR on NAFLD was evaluated in C57BL/6\u202fJ mice on high diet (HFD). The effect of BBR was also investigated in HepG2 and AML12\u202fcells exposed to high glucose and . Oil red O staining was performed to detect triglyceride (TG) level. Quantitative real-time polymerase chain reaction and Western blot were used to detect the messenger ribonucleic (mRNA) and protein expression of target genes. The activity of SCD1 promoter was measured by dual-luciferase reporter assay.The expression of SCD1 was increased in the liver of NAFLD patients and ob/ob mice. BBR reduced hepatic TG accumulation and decreased the expressions of hepatic SCD1 and other TG synthesis related genes both in vivo and in vitro. Knockdown of SCD1 expression mimicked the effect of BBR decreasing TG level in steatotic hepatocytes, whereas overexpression of SCD1 attenuated the effect of BBR. Mechanistically, BBR promoted the phosphorylation of AMP-activated protein kinase (AMPK) and sterol regulatory element-binding protein-1c (SREBP-1c) in HepG2 cells and the liver of HFD-fed mice. Activation of the AMPK-SREBP-1c pathway and sterol regulatory element (SRE) motif in SCD1 promoter (-920/-550) was responsible for the BBR-induced suppression of SCD1.BBR reduces liver TG synthesis and attenuates hepatic steatosis through the activation of AMPK-SREBP-1c-SCD1 pathway.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Hepatoprotective and Antioxidant Activities of Oil from Baru Almonds ( Vog.) in a Preclinical Model of Lipotoxicity and Dyslipidemia.

The oil obtained from baru ( Vog.) almonds exhibits high energy value and is reported in popular medicine for the treatment of rheumatic diseases and reproductive disturbances. Although baru oil is used in domestic cuisine, the chemical characterization of this oil and its effects on are still poorly understood. Therefore, this study evaluated the fatty (FA) profile and the effects of baru oil on liver and aorta in a murine model of dyslipidemia. The chromatographic profile of baru oil showed high levels of unsaturated FAs, especially oleic . Saturated FAs, such as and lignoceric acids, were found in lower amounts. Hypercholesterolemia was induced in male Wistar rats by daily administration of a emulsion by gavage for 15 weeks. Biochemical and histopathological analysis were performed on serum, aorta, and liver. The results demonstrated that animals developed marked hypercholesterolemia, liver steatosis, and increased peroxidation in the aorta. Treatment with baru oil attenuated peroxidation and drastically reduced liver damage, especially ballooning degeneration and steatosis. By restricting vascular and hepatic injury, this oil showed potential applicability as a functional food, reinforcing its use in popular medicine and domestic cuisine.

Keyword: fat metabolism

Reduces the Autophagic Flux and Insulin Sensitivity Through the Activation of the Free Fatty Receptor 1 (FFAR1) in the Hypothalamic Neuronal Cell Line N43/5.

Chronic consumption of high diets (HFDs), rich in saturated fatty acids (SatFAs) like (PA), is associated with the development of obesity and obesity-related metabolic diseases such as type II diabetes mellitus (T2DM). Previous studies indicate that PA accumulates in the hypothalamus following consumption of HFDs; in addition, HFDs consumption inhibits autophagy and reduces insulin sensitivity. Whether malfunction of autophagy specifically in hypothalamic neurons decreases insulin sensitivity remains unknown. PA does activate the Free Fatty Receptor 1 (FFAR1), also known as G protein-coupled receptor 40 (GPR40); however, whether FFAR1 mediates the effects of PA on hypothalamic autophagy and insulin sensitivity has not been shown. Here, we demonstrate that exposure to PA inhibits the autophagic flux and reduces insulin sensitivity in a cellular model of hypothalamic neurons (N43/5 cells). Furthermore, we show that inhibition of autophagy and the autophagic flux reduces insulin sensitivity in hypothalamic neuronal cells. Interestingly, the inhibition of the autophagic flux, and the reduction in insulin sensitivity are prevented by pharmacological inhibition of FFAR1. Our findings show that dysregulation of autophagy reduces insulin sensitivity in hypothalamic neuronal cells. In addition, our data suggest FFAR1 mediates the ability of PA to inhibit autophagic flux and reduce insulin sensitivity in hypothalamic neuronal cells. These results reveal a novel cellular mechanism linking PA-rich diets to decreased insulin sensitivity in the hypothalamus and suggest that hypothalamic autophagy might represent a target for future T2DM therapies.

Keyword: fat metabolism

Fibroblast Growth Factor 21 Stimulates Pancreatic Islet Autophagy via Inhibition of AMPK-mTOR Signaling.

Islet autophagy plays a role in glucose/ in type 2 diabetes mellitus. Meanwhile, fibroblast growth factor 21 (FGF21) has been found to regulate insulin sensitivity and glucose homeostasis. Whether FGF21 induces islet autophagy, remains to be elucidated. This study aimed to explore the physiological roles and signaling pathways involved in FGF21-stimulated islet autophagy under glucolipotoxic conditions.C57/BL6J mice were fed a standard diet or high- diet (HFD) for 12 weeks, and islets were isolated from normal and knockout (KO) mice. Isolated islets and INS-1E cells were exposed to normal and high-concentration glucose and with/without FGF21 or AMPK inhibitor compound C. Real-time PCR, Western blot and immunohistochemistry/transmission electron microscopy were performed for the expression of targeted genes/proteins.HFD-treated mice showed increases in fasting plasma glucose, body weight and impaired glucose tolerance; islet protein expression of FGF21 was induced after HFD treatment. Protein expression levels of FGF21 and LC3-II (autophagy marker) were induced in mouse islets treated with high concentrations of and glucose, while phosphorylation of AMPK was reduced, compared with controls. In addition, induction of LC3-II protein expression was reduced in islets isolated from KO mice. Furthermore, exogenous administration of FGF21 diminished phosphorylation of AMPK and stimulated protein expression of LC3-II. Consistently, compound C significantly induced increased expression of LC3-II protein.Our data indicate that glucolipotoxicity-induced FGF21 activation mediates islet autophagy via AMPK inhibition, and further consolidate the evidence for the FGF21/analog being a pharmacotherapeutic target for obesity and its related T2DM.

Keyword: fat metabolism

Gut Microbiota and Metabolome Response of Seed Oil on Disorder Induced by Excess Alcohol Consumption.

This study investigated the modulatory effects of seed oil (DISO), which was rich in palmitoleic (55.25%), (12.25%), and oleic (28.74%), on alcohol-induced disorder in mice. Fifty mice were orally administered with 38% alcohol (0.4 mL/day) and without or with DISO (3, 6, and 12 g/kg) for consecutive 12 weeks. DISO inhibited the alcohol-induced weight loss and liver function abnormality ( < 0.01) and shifted the profiles of cecal microbiome: elevating the abundance of , __004 ( < 0.05) and decreasing abundance of ( < 0.05). This treatment also regulated metabolome response of amino and in cecal content: upregulating 5-hydroxyindole-3-acetic ( < 0.05), 6-hydroxynicotinic , 5-methoxytryptamine, nicotinamide, and nicotinic ( < 0.1) and downregulating androsterone, tryptophan, and indole-3-acetamide ( < 0.05). DISO protected against alcoholic liver injury and gut microbiota dysbiosis by enriching the relative abundance of , which was positively associated with the improvement of intestinal permeability and tryptophan .

Keyword: fat metabolism

mTORC2 Regulates Lipogenic Gene Expression through PPAR to Control Synthesis in Bovine Mammary Epithelial Cells.

The mechanistic target of rapamycin complex 2 (mTORC2) primarily functions as an effector of insulin/PI3K signaling to regulate cell proliferation and is associated with cell . However, the function of mTORC2 in is not well understood. In the present study, mTORC2 was inactivated by the ATP-competitive mTOR inhibitor AZD8055 or shRNA targeting in primary bovine mammary epithelial cells (pBMECs). MTT assay was performed to examine the effect of AZD8055 on cell proliferation. ELISA assay and GC-MS analysis were used to determine the content of . The mRNA and protein expression levels were investigated by RT/real-time PCR and western blot analysis, respectively. We found that cell proliferation, mTORC2 activation, and secretion were inhibited by AZD8055. was knocked down and mTORC2 activation was specifically attenuated by the shRNA. Compared to control cells, the expression of the transcription factor gene and the lipogenic genes , , , and was downregulated in silencing cells. As a result, the content of intracellular triacylglycerol (TAG), (PA), docosahexaenoic (DHA), and other 16 types of fatty was decreased in the treated cells; the accumulation of TAG, PA, and DHA in cell culture medium was also reduced. Overall, mTORC2 plays a critical role in regulating lipogenic gene expression, synthesis, and secretion in pBMECs, and this process probably is through PPAR. This finding provides a model by which lipogenesis is regulated in pBMECs.

Keyword: fat metabolism

Dietary modulation of energy homoeostasis and metabolic-inflammation.

Dietary intake and nutritional status is an important environmental factor which can modulate metabolic-inflammation. In recent years, research has made significant advances in terms of understanding the impact of dietary components on metabolic-inflammation, within the context of obesity, type-2 diabetes (T2D) and CVD risk. Our work demonstrated that different fatty acids differentially modulate metabolic-inflammation, initially focusing on Nod-like receptor family, pyrin domain-containing three protein (NLRP3) inflammasome mediated IL-1β biology and insulin signalling. However, the paradigm is more complex, wherein data from the immunology field clearly show that nature of cellular energy is a key determinant of inflammation. Whilst metabolic-inflammation is a critical biological interaction, there is a paucity of data in relation to the nature and the extent to which nutritional status affects metabolic-inflammation. The complex paradigm will be discussed within the context of if/how dietary components, in particular fatty acids, may modulate obesity, T2D and CVD risk, via inflammatory and metabolic processes.

Keyword: fat metabolism

Influence of nutrient formulations on growth, yield, carbon partitioning and biodiesel quality potential of Botryococcus sp. and Chlorella sp.

The study was conducted to analyse the influence of three nutrient formulations, namely BG-11 medium, BBM and TAP medium, on growth potential and yield of two microalgal genera (Botryococcus sp. and Chlorella sp.) and to study the roles of N, P and other major nutrients. The study focussed on the general patterns of starch and synthesis and storage and to further assess how photosynthetic carbon partitioning into starch and is altered by conditions in growth media such as N and C presence as seen in BG11 medium which are known to induce neutral production and the lack of it in BBM and TAP medium. BG-11 medium performed better as compared to BBM and TAP medium in terms of biomass productivity and yield. The yield was highest in Botryococcus sp. (63.03% dry wt.) and Chlorella sp. (50.27% dry wt.) at 30th day of incubation. Mean biomass productivity was highest for Botryococcus in BBM medium (6.14\xa0mg/L/day) and for Chlorella in BG-11 medium (4.97\xa0mg/L/day). Mean productivity (50.78% and 39.36%) was highest in BG11 medium for both Botryococcus and Chlorella species, respectively. A sharp decline in sugar content was observed in the late stationary phase of growth from 30th day to 45th day. Fatty methyl ester (FAME) profile of the extracted lipids showed predominantly oleic , followed by and stearic in both the strains when grown in BG-11 medium. The other biodiesel quality parameters were in accordance with the international standards. A complex relationship was found between chemical composition and biodiesel properties. Proximity analysis indicated that the fuel properties of biodiesels are determined by a number of parameters and by the combination of different chemical compositions. The results provide an insight into organic carbon partitioning into compounds and how the organism\'s changes due to N-deplete culturing in TAP medium and inorganic carbon source availability as seen in BG-11 and BBM medium.

Keyword: fat metabolism

Obesity Mediates Apoptosis and Extracellular Matrix Metabolic Imbalances via MAPK Pathway Activation in Intervertebral Disk Degeneration.

Obesity may promote intervertebral disc degeneration (IDD) by non-mechanical means, by influencing levels of free fatty acids which could impair cell . This study aims to establish metabolic factors in obesity-related IDD independent of mechanical loading. In clinical study, we retrospectively reviewed 128 volunteers (73 males, 55 females, aged 29-88 years) and compared their grades of disk degeneration with obesity-related factors such as body weight, BMI, and serum levels. Clinically, the IDD group showed increased age, BMI and serum triglyceride. Triglyceride was a significant risk factor for IDD even after correction for BMI and age (P = 0.007). In obesity animal model, rats were fed a high- diet (HFD) in order to study its effects on disk and apoptosis. HFD rats had significantly higher serum levels of lipids, including triglyceride and non-esterified fatty , and showed significantly decreased markers of anabolism, increased catabolism and apoptosis in disk. Finally, rat nucleus pulposus (NP) cells were stimulated with a fatty (, PA) to gauge its effects on cell and apoptosis. Cell culture studies showed that NP cells exposed to PA showed increased apoptosis for activation of caspase 3, 7, 9, and PARP, which was primarily via the MAPK signal pathway, especially ERK pathway. In conclusion, hypertriglyceridemia can lead to IDD, independently of age and BMI. Hypertriglyceridemia appears to mediate disk cell apoptosis and matrix catabolism primarily via the ERK pathway.Copyright © 2019 Zhang, Chen, Huang, Wang, Shan, Liu, Chen, Li, Fan and Zhao.

Keyword: fat metabolism

Pex11a deficiency causes dyslipidaemia and obesity in mice.

Peroxisomes play a central role in . We previously demonstrated that Pex11a deficiency impairs peroxisome abundance and fatty β-oxidation and results in hepatic triglyceride accumulation. The role of Pex11a in dyslipidaemia and obesity is investigated here with Pex11a knockout mice (Pex11a ). Metabolic phenotypes including tissue weight, glucose tolerance, insulin sensitivity, cholesterol levels, fatty profile, oxygen consumption, physical activity were assessed in wild-type (WT) and Pex11a fed with a high- diet. Molecular changes and peroxisome abundance in adipose tissue were evaluated through qRT-PCR, Western blotting, and Immunofluorescence. Pex11a showed increased mass, decreased skeletal muscle, higher cholesterol levels, and more severely impaired glucose and insulin tolerance. Pex11a consumed less oxygen, indicating a decrease in fatty oxidation, which is consistent with the accumulation of very long- and long-chain fatty acids. Adipose (C16:0) levels were elevated in Pex11a , which may be because of dramatically increased fatty synthase mRNA and protein levels. Furthermore, Pex11a deficiency increased ventricle size and macrophage infiltration, which are related to the reduced physical activity. These data demonstrate that Pex11a deficiency impairs physical activity and energy expenditure, decreases fatty β-oxidation, increases de novo lipogenesis and results in dyslipidaemia and obesity.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fat metabolism

Amelioration of Endoplasmic Reticulum Stress by Mesenchymal Stem Cells via Hepatocyte Growth Factor/c-Met Signaling in Obesity-Associated Kidney Injury.

Recent advances in the understanding of suggest a critical role of endoplasmic reticulum (ER) stress in obesity-induced kidney injury. Hepatocyte growth factor (HGF) is a pleiotropic cytokine frequently featured in stem cell therapy with distinct renotropic benefits. This study aims to define the potential link between human induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs)/bone marrow-derived MSCs (BM-MSCs) and ER stress in lipotoxic kidney injury induced by (PA) in renal tubular cells and by high- diet (HFD) in mice. iPS-MSCs or BM-MSCs alleviated ER stress (by preventing induction of Bip, chop, and unfolded protein response), inflammation (Il6, Cxcl1, and Cxcl2), and apoptosis (Bax/Bcl2 and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells) in renal cortex of animals exposed to HFD thus mitigating histologic damage and albuminuria, via activating HGF/c-Met paracrine signaling that resulted in enhanced HGF secretion in the glomerular compartment and c-Met expression in the tubules. Coculture experiments identified glomerular endothelial cells (GECs) to be the exclusive source of glomerular HGF when incubated with either iPS-MSCs or BM-MSCs in the presence of PA. Furthermore, both GEC-derived HGF and exogenous recombinant HGF attenuated PA-induced ER stress in cultured tubular cells, and this effect was abrogated by a neutralizing anti-HGF antibody. Taken together, this study is the first to demonstrate that MSCs ameliorate lipotoxic kidney injury via a novel microenvironment-dependent paracrine HGF/c-Met signaling mechanism to suppress ER stress and its downstream pro-inflammatory and pro-apoptotic consequences. Stem Cells Translational Medicine 2019;8:898&910.© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

Keyword: fat metabolism

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic fatty liver disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high--diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human liver cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the liver than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

Keyword: fat metabolism

Adipose tissue dysfunction is associated with low levels of the novel Hydroxystearic Acids.

Adipose tissue dysfunction is considered an important contributor to systemic insulin resistance and Type 2 diabetes (T2D). Recently, a novel family of endogenous lipids, hydroxy stearic acids (PAHSAs), was discovered. These have anti-diabetic and anti-inflammatory effects in mice and are reduced in serum and adipose tissue of insulin resistant humans. In the present study, we investigate if adipose tissue dysfunction is associated with reduced PAHSA levels in human subjects and if PAHSAs influence adipocyte differentiation. Our results show that low expression of adipocyte GLUT4 and adipocyte hypertrophy, markers of adipose tissue dysfunction, are associated with reduced expression of key enzymes for de novo lipogenesis and adipose tissue levels of PAHSAs in human subjects. We also show that GLUT4 is not only a marker of adipose tissue dysfunction, but may be causally related to the observed impairments. PAHSAs may also act locally in the adipose tissue to improve adipogenesis through a mechanism bypassing direct activation of peroxisome proliferator-activated receptor (PPARγ). The discovery of PAHSAs and our current results provide novel insights into positive effects of species in adipose tissue and mechanisms by which dysfunctional adipose tissue is associated with insulin resistance and risk of developing T2D.

Keyword: fat metabolism

Polydatin attenuates diet-induced nonalcoholic steatohepatitis and fibrosis in mice.

Non-alcoholic steatohepatitis (NASH) is characterized by accumulation in hepatocytes and inflammatory cell infiltration. In view of the anti-oxidative and anti-inflammatory effects of polydatin, the current study aimed to investigate the pharmacological effects of polydatin on NASH and its related fibrosis. C57BL/6 mice were fed with methionine-choline deficient (MCD) diet to induce NASH and liver fibrosis, and treated with or without polydatin (5 mg/kg, every other day, i.p) for 4 weeks. HepG2 cells induced by (PA) were treated with polydatin. The elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), active caspase-3, TUNEL-positive cells, and triglyceride content were decreased by polydatin treatment. In addition, administration of polydatin to MCD-fed mice reduced oxidative stress by down-regulating NOX4 enzymes. Furthermore, the reduction in inflammation and CD68 macrophage activation correlated with inhibition of toll-like receptor (TLR)-4/NF-κB p65 signaling pathway by polydatin treatment. Polydatin also attenuated accumulation, inflammation and apoptosis in HepG2 cells challenged by (PA) combined with or without lipopolysaccharide (LPS). Finally, the reduction of hepatic fibrosis by polydatin treatment corresponded to a reduction in hepatic gene expression of fibrosis markers. These results suggest that polydatin prevents NASH and fibrosis via inhibition of oxidative stress and inflammation, highlighting polydatin as a potential therapeutic agent for prevention and treatment of NASH.

Keyword: fat metabolism

Free fatty -induced histone acetyltransferase activity accelerates accumulation in HepG2 cells.

Non-alcoholic fatty liver disease (NAFLD) is a common metabolic disease triggered by epigenetic alterations, including lysine acetylation at histone or non-histone proteins, affecting the stability or transcription of lipogenic genes. Although various natural dietary compounds have anti-lipogenic effects, their effects on the acetylation status and in the liver have not been thoroughly investigated.Following oleic- (OPA)-induced accumulation in HepG2 cells, the acetylation status of histone and non-histone proteins, HAT activity, and mRNA expression of representative lipogenic genes, including γ, , , and , were evaluated. Furthermore, correlations between accumulation and HAT activity for 22 representative natural food extracts (NExs) were evaluated.Non-histone protein acetylation increased following OPA treatment and the acetylation of histones H3K9, H4K8, and H4K16 was accelerated, accompanied by an increase in HAT activity. OPA-induced increases in the mRNA expression of lipogenic genes were down-regulated by C-646, a p300/CBP-specific inhibitor. Finally, we detected a positive correlation between HAT activity and accumulation (Pearson\'s correlation coefficient = 0.604) using 22 NExs.Our results suggest that NExs have novel applications as nutraceutical agents with HAT inhibitor activity for the prevention and treatment of NAFLD.

Keyword: fat metabolism

Chronic Heat Stress Induces Acute Phase Responses and Serum Metabolome Changes in Finishing Pigs.

Heat stress (HS) is a main environmental challenge affecting the animal welfare and production efficiency in pig industry. In recent years, numerous reports have studied the alterations in gene expressions and protein profiles in heat-stressed pigs. However, the use of metabolome to unravel adaptive mechanisms of finishing pig in response to chronic HS have not yet been elucidated. We aimed to investigate the effects of chronic HS on serum metabolome in finishing pigs, and to identify the biomarkers of heat stress. Pigs (n = 8 per treatment) were exposed to either thermal neutral (TN; 22 °C) or heat stress (HS, 30 °C) conditions for three weeks. Serum metabonomics of TN- and HS-treated pigs were compared using the GC-MS approach. Metabonomics analysis revealed that twenty-four metabolites had significantly different levels in TN compared to HS (variable importance in the projection values >1 and < 0.05). These metabolites are involved in carbohydrate, amino , fatty , amines , and gut microbiome-derived . Three serum monoses (glucose, mannose 2, and galactose) and 6-phosphogluconic were decreased, indicating insufficient source of fuel for energy supply, resulting in negative energy balance (NEB) in heat-stressed pigs. Increased levels of non-esterified fatty (myristic , , and linoleic ) and short-chain fatty acids (3-hydroxybutanoic and maleic ) suggested decomposition compensating for energy shortage, which was an adaptive response to NEB. Increased concentrations of fluorine, lyxose 1, and D-galacturonic were significantly correlated with the levels of acute phase proteins (HP, LBP, α2-HSG, and Lysozyme), suggesting acute phase response in HS-stressed pigs. These metabolites are expected to be novel biomarkers of chronic HS in pigs, yet the use of which awaits further validation.

Keyword: fat metabolism

Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Droplets in Cultured Hepatocytes.

A fluorescence microscopic method for characterizing size, quantity, and oxidation of droplets (LDs) in HepG2 cells was developed. LDs were induced by (PA), oleic (OA), or linoleic acids (LA) and stained with two fluorescent probes for neutral lipids and peroxides. Each fatty increased the number of LDs and oxidized LDs (oxLDs) and the degree of LD oxidation time dependently, as well as increased intracellular triglyceride hydroperoxides. LDs induced by LA without 2,2\'-azobis(2-amidinopropane)dihydrochloride (AAPH) showed the most significant oxidation degree over PA and OA, especially in large LDs (area ≥ 3 μm, oxLD/LD = 52.3 ± 21.7%). Under this condition, two food-derived antioxidants were evaluated, and both of them significantly improved the LD characteristics. Moreover, chlorogenic reduced the quantity of large LDs by 74.0-87.6% in a dose-dependent manner. The proposed method provides a new approach to evaluate the effect of dietary antioxidants on LD characteristics.

Keyword: fat metabolism

of 13C-Labeled Fatty Acids in Term Human Placental Explants by Liquid Chromatography-Mass Spectrometry.

Placental transport and are poorly understood despite the importance for fetal development and lifelong health. We aimed to explore fatty (FA) processing in human villous placental explants from seven uncomplicated term singleton pregnancies delivered by elective cesarean section. Explants were treated with stable isotope-labeled (13C-PA), oleic (13C-OA), or docosahexaenoic (13C-DHA) for 3, 24, or 48 hours. Stable isotope-labeled lipids synthesized by placental explants from labeled FA were quantified, alongside endogenous unlabeled placental lipids, by liquid chromatography-mass spectrometry. Labeled phosphatidylcholines (PCs), triacylglycerols (TAGs), and phosphatidylethanolamines were detected in explants, whereas labeled lysophosphatidylcholines were found in both explants and conditioned media. 13C-PA was primarily directed into PC synthesis (74% of 13C-PA-labeled lipids), whereas 13C-OA was directed almost equally into PC and TAG synthesis (45% and 53%, respectively, of 13C-OA-labeled lipids). 13C-DHA was only detectable in TAGs. TAGs demonstrated the highest isotopic enrichment for all 13C-FAs with 13C-OA-TAGs comprising >50% of total OA-TAGs (unlabeled and labeled), consistent with TAGs being a labile and accessible reservoir for FA storage. Variations in incorporation were correlated to maternal glycemia and body mass index, suggesting that this experimental model could be used to investigate the effect of maternal factors on placental . We conclude that metabolic partitioning of freshly imported FAs into labile and less labile reservoirs in placenta is FA dependent. This process may partly mediate the physiological preferential transplacental transfer of particular FAs to the fetus, but may also be implicated in the fetoplacental pathophysiology of maternal metabolic dysfunction.Copyright © 2019 Endocrine Society.

Keyword: fat metabolism

Triterpenoids from Hibiscus sabdariffa L.\xa0with PPARδ/γ Dual Agonist Action: In Vivo, In Vitro and In Silico Studies.

is a medicinal plant consumed as a diuretic and anti-obesity remedy. Several pharmacological studies have shown its beneficial effects in . Peroxisome proliferator-activated receptors and may play a role in the actions of These nuclear receptors regulate and glucose and are therapeutic targets for type 2 diabetes. This research aimed to perform a phytochemical study guided by a bioassay from to identify compounds with peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor agonist activity, supported by messenger ribonucleic expression, molecular docking, accumulation, and an antihyperglycemic effect. An oral glucose tolerance test in mice with the aqueous extract of and the dichloromethane extract of was performed. The dichloromethane extract of exhibited an antihyperglycemic effect. The dichloromethane extract of was fractioned, and four fractions were evaluated in 3T3-L1 adipocytes on peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 messenger ribonucleic expression. Fraction F3 exhibited peroxisome proliferator-activated receptor / dual agonist activity, and a further fractionation yielded two subfractions, F3-1 and F3-2, which also increased peroxisome proliferator-activated receptor and peroxisome proliferator-activated receptor expression. Subfractions were analyzed by GC/MS. The main compounds identified in F3-1 were linoleic , oleic , and , while in F3-2, the main compounds identified were -amyrin and lupeol. These molecules were subjected to molecular docking analysis. -Amyrin and lupeol showed the highest affinity. Moreover, both produced an increase in peroxisome proliferator-activated receptor , peroxisome proliferator-activated receptor , fatty transporter protein, and glucose transporter type 4 expression. Additionally, -amyrin and lupeol decreased accumulation in 3T3-L1 adipocytes and blood glucose in mice. Until now, -amyrin and lupeol have not been reported with activity on peroxisome proliferator-activated receptors. This study provides evidence that -amyrin and lupeol possess antidiabetic effects through a peroxisome proliferator-activated receptor / dual agonist action.Georg Thieme Verlag KG Stuttgart · New York.

Keyword: fat metabolism

The effect of enterolactone on sphingolipid pathway and hepatic insulin resistance development in HepG2 cells.

Obesity and type 2 diabetes mellitus, correlate with increased tissue concentration of sphingolipids, which directly interfere with insulin signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid and decrease hepatic insulin resistance development in a overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and insulin signaling pathway proteins and glucose regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of insulin-stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher fatty acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Bioconversion of barley straw lignin into biodiesel using Rhodococcus sp. YHY01.

Rhodococcus sp. YHY01 was studied to utilize various lignin derived aromatic compounds. It was able to utilize p-coumaric , cresol, and 2,6 dimethoxyphenol and resulted in biomass production i.e. 0.38\u202fg dcw/L, 0.25\u202fg dcw/L and 0.1\u202fg dcw/L, and accumulation i.e. 49%, 40%, 30%, respectively. The half maximal inhibitory concentration (IC) value for p-coumaric (13.4\u202fmM), cresol (7.9\u202fmM), and 2,6 dimethoxyphenol (3.4\u202fmM) was analyzed. Dimethyl sulfoxide (DMSO) solubilized barley straw lignin fraction was used as a carbon source for Rhodococcus sp. YHY01 and resulted in 0.130\u202fg dcw/L with 39% w/w accumulation. Major fatty acids were (C16:0) 51.87%, palmitoleic (C16:l) 14.90%, and oleic (C18:1) 13.76%, respectively. Properties of biodiesel produced from barley straw lignin were as iodine value (IV) 27.25, cetane number (CN) 65.57, cold filter plugging point (CFPP) 14.36, viscosity (υ) 3.81, and density (ρ) 0.86.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

Silibinin ameliorates hepatic accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis (NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of fatty acids in liver to relieve accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: fat metabolism

Furanoid F- F6 Uniquely Induces NETosis Compared to C16 and C18 Fatty Acids in Human Neutrophils.

Various biomolecules induce neutrophil extracellular trap (NET) formation or NETosis. However, the effect of fatty acids on NETosis has not been clearly established. In this study, we focused on the NETosis-inducing ability of several molecules. We extracted the molecules present in Arabian Gulf catfish (, Val) skin gel, which has multiple therapeutic activities. Gas chromatography⁻mass spectrometry (GC-MS) analysis of the fraction-3 from the gel with NETosis-inducing activity contained fatty acids including a furanoid F- (F6; 12,15-epoxy-13,14-dimethyleicosa-12,14-dienoic ) and common long-chain fatty acids such as (PA; C16:0), palmitoleic (PO; C16:1), stearic (SA; C18:0), and oleic (OA; C18:1). Using pure molecules, we show that all of these fatty acids induce NETosis to different degrees in a dose-dependent fashion. Notably, F6 induces a unique form of NETosis that is rapid and induces reactive oxygen species (ROS) production by both NADPH oxidase (NOX) and mitochondria. F6 also induces citrullination of histone. By contrast, the common fatty acids (PA, PO, SA, and OA) only induce NOX-dependent NETosis. The activation of the kinases such as ERK (extracellular signal-regulated kinase) and JNK (c-Jun N-terminal kinase) is important for long-chain fatty -induced NETosis, whereas, in F--induced NETosis, Akt is additionally needed. Nevertheless, NETosis induced by all of these compounds requires the final chromatin decondensation step of transcriptional firing. These findings are useful for understanding F-- and other fatty -induced NETosis and to establish the active ingredients with therapeutic potential for regulating diseases involving NET formation.

Keyword: fat metabolism

Genomics of -laden human hepatocyte cultures enables drug target screening for the treatment of non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) is a major health burden in need for new medication. To identify potential drug targets a genomic study was performed in -laden primary human hepatocyte (PHH) and human hepatoma cell cultures.PHH, HuH7 and HepG2 hepatoma cell cultures were treated with lipids and/or TNFα. Intracellular load was quantified with the ORO assay. The Affymetrix HG-U133+ array system was employed to perform transcriptome analysis. The droplet (LD) growth and fusion was determined by fluorescence microscopy. LD associated proteins were imaged by confocal immunofluorescence microscopy and confirmed by Western immunoblotting. Bioinformatics defined perturbed metabolic pathways.Whole genome expression profiling identified 227, 1031 and 571 significant regulated genes. Likewise, the combined and TNFα treatment of PHH, HuH7 and HepG2 cell cultures revealed 154, 1238 and 278 differentially expressed genes. Although genomic responses differed among in-vitro systems, commonalities were ascertained by filtering the data for LD associated gene regulations. Among others the LD-growth and fusion associated cell death inducing DFFA like effector C (CIDEC), perilipins (PLIN2, PLIN3), the synaptosome-associated-protein 23 and the vesicle associated membrane protein 3 were strongly up-regulated. Likewise, the PPAR targets pyruvate-dehydrogenase-kinase-4 and angiopoietin-like-4 were up-regulated as was hypoxia-inducible droplet-associated (HILPDA), flotilin and FGF21. Their inhibition ameliorates triglyceride and cholesterol accumulation. TNFα treatment elicited strong induction of the chemokine CXCL8, the kinases MAP3K8, MAP4K4 and negative regulators of cytokine signaling, i.e. SOCS2&SOCS3. Live cell imaging of DsRED calreticulin plasmid transfected HuH7 cells permitted an assessment of LD growth and fusion and confocal immunofluorescence microscopy evidenced induced LD-associated PLIN2, CIDEC, HIF1α, HILPDA, JAK1, PDK4 and ROCK2 expression. Notwithstanding, CPT1A protein was repressed to protect mitochondria from overload. Pharmacological inhibition of the GTPase-dynamin and the fatty transporter-2 reduced uptake by 28.5 and 35%, respectively. Finally, a comparisons of in-vitro/NAFLD patient biopsy findings confirmed common gene regulations thus demonstrating clinical relevance.The genomics of -laden hepatocytes revealed LD-associated gene regulations and perturbed metabolic pathways. Immunofluorescence microscopy confirmed expression of coded proteins to provide a rationale for therapeutic intervention strategies. Collectively, the in-vitro system permits testing of drug candidates.

Keyword: fat metabolism

Transcriptome analysis of elaiosomes and seeds provide insights into fatty biosynthesis.

Elaiosomes are specialized fleshy and edible seed appendages dispersed by ants. Lipids are the primary components of elaiosomes. is a well-known plant, the seeds of which are dispersed by ants. Previous studies have identified the presence of primary fatty acids in its elaiosomes and seeds. However, the molecular mechanisms underlying fatty biosynthesis in elaiosomes remain unknown.In order to gain a comprehensive transcriptional profile of the elaiosomes and seeds of , and understand the expression patterns of genes associated with fatty biosynthesis, four different developmental stages, including the flower-bud (Ch01), flowering (Ch02), young seed (Ch03), and mature seed (Ch04) stages, were chosen to perform whole-transcriptome profiling through the RNA-seq technology (Illumina NGS sequencing).A total of 63,064 unigenes were generated from 12 libraries. Of these, 7,323, 258, and 11,540 unigenes were annotated with 25 Cluster of Orthologous Groups, 43 Gene Ontology terms, and 373 Kyoto Encyclopedia of Genes and Genomes pathways, respectively. In addition, 322 genes were involved in transport and , and 508 genes were involved in the pathways. A total of 41 significantly differentially expressed genes (DEGs) involved in the pathways were identified, most of which were upregulated in Ch03 compared to Ch02, indicating that fatty biosynthesis primarily occurs during the flowering to the young seed stages. Of the DEGs, acyl-ACP thioesterases, acyl carrier protein desaturase (), and malonyl CoA-ACP transacylase were involved in synthesis; stearoyl-CoA desaturase and were involved in oleic synthesis, and acyl- omega-6 desaturase was involved in linoleic synthesis.

Keyword: fat metabolism

Downregulates Thyroglobulin (Tg), Sodium Iodide Symporter (NIS), and Thyroperoxidase (TPO) in Human Primary Thyrocytes: A Potential Mechanism by Which Lipotoxicity Affects Thyroid?

Our previous studies suggested that the thyroid might be a target organ affected by lipotoxicity, which might be partially related to the increasing prevalence of subclinical hypothyroidism. However, the underlying molecular mechanism is not yet clearly established. This study aimed to assess the effect of stimulation on thyrocyte function. Upon stimulation, intracellular contents of lipids, as well as the expression and activity of three key molecules in thyroid hormone synthesis (i.e., thyroglobulin, sodium iodide symporter, and thyroperoxidase), were determined in human primary thyrocytes. The contents of BODIPY® FL C16 (the fluorescently labeled analogue) entering into the thyrocytes were gradually increased with time extending. Accordingly, the intracellular accumulation of both triglyceride and free fatty acids increased in dose- and time-dependent manners. The effect of stimulation on thyroid hormone synthesis was then determined. Both the mRNA and protein levels of thyroglobulin, sodium iodide symporter, and thyroperoxidase were decreased following stimulation. Consistently, upon stimulation, the secreted thyroglobulin levels in supernatants, I uptake, and extracellular thyroperoxidase activity were all decreased in a dose-dependent manner. Our results demonstrated that upon stimulation, the expressions of the key molecules (thyroglobulin, sodium iodide symporter, and thyroperoxidase) were reduced and their activities were suppressed, which might lead to impaired thyroid hormone synthesis.

Keyword: fat metabolism

PAQR3 regulates phosphorylation of FoxO1 in insulin-resistant HepG2 cells via NF-κB signaling pathway.

Insulin resistance is a significant feature of type 2 diabetes mellitus and glucose and disorders. Activation of NF-κB signaling pathway plays an important role in the formation of insulin resistance. FoxO1 plays a major role in regulating glucose and , as well as insulin signaling pathway. Previous studies have shown that Progestin and AdipoQ Receptor 3 (PAQR3) suppresses the activity of PI3K/Akt, which is an upstream pathway of FoxO1, and additionally promotes the pathological process of diabetic renal inflammatory fibrosis via activating NF-κB pathway. On this basis, it has caused us great concern whether NF-κB is involved in PAQR3 regulation of FoxO1 under insulin resistance. In this study, we aimed to investigate whether PAQR3 regulates phosphorylation of FoxO1 via NF-κB pathway in (PA)-induced insulin-resistant HepG2 cells, thereby causing glucose and disorders. We found that PA stimulation and PAQR3 overexpression decreased the phosphorylation of FoxO1 and the expressions of glucokinase (GCK) and low density lipoprotein receptor (LDLR), in addition, promoted the nuclear accumulation of NF-κB. Inhibition of NF-κB pathway increased the phosphorylation of FoxO1 and the expressions of GCK and LDLR which were downregulated by PA stimulation and PAQR3 overexpression. Taken together, in PA-induced insulin-resistant HepG2 cells, PAQR3 might regulate the phosphorylation of FoxO1 and the expressions of GCK and LDLR through NF-κB pathway, thereby regulating the glucose and disorders induced by insulin resistance.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

The Differentiation of Spinal Cord Motor Neurons is Associated with Changes of the Mitochondrial Phospholipid Cardiolipin.

Motor neuron damage caused by diseases, traumatic insults or de-afferentation of the spinal cord is often incurable due to the poor intrinsic regenerative capacity. Moreover, regenerated peripheral nerves often do not reach normal functionality. Here, we investigated cardiolipin in the process of neuro-differentiation, since cardiolipin is closely linked to the mitochondrial energy supply in cells. The NSC-34 hybrid cell line, produced by fusing neuroblastoma cells with primary spinal cord motor neurons, was used, since it shares several morphological and physiological characteristics with mature primary motor neurons. Their neuro-differentiation was supported by switching from normal to differentiation medium or by fatty supplementation. Differentiation was evaluated by measuring neurite-sprouting parameters and PPARα expression. Cellular fatty distribution was analyzed to indicate changes in during differentiation. Cardiolipin was characterized by acyl-chain composition and the distribution of molecular cardiolipin species. Both, the switch from normal to differentiation medium as well as the administration of and oleic promoted neuro-differentiation. Stimulated differentiation was accompanied by changes in cardiolipin content and composition. The positive correlation between neuro-differentiation and concentration of those molecular cardiolipin species containing and oleic implied a link between differentiation of NSC-34 cells and cardiolipin . We further demonstrated the impact of cellular , and particularly cardiolipin , during and NSC-34 neuritogenesis. Thus, cardiolipin may represent a new therapeutic target for axon regeneration after peripheral nerve injuries or when axon sprouting is required to compensate for motor neuron loss in response to aging and/or disease.Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.

Keyword: fat metabolism

-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic β-cells.

Free fatty receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic β-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated insulin secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of β-cell function. FFAR1 plays a key role in the development of type 2 diabetes (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of β-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic β-cells. Key players of the insulin signaling pathway, such as mTOR, Akt, IRS-1, and the insulin receptor (INSR1β), were selected as candidates to be analyzed under lipotoxic conditions.We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1β and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1β mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus.In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in β-cells under lipotoxic conditions.

Keyword: fat metabolism

Uncommon properties of biosynthesis of isolated plastids in the unicellular red alga .

Red algae are a large group of photosynthetic eukaryotes that diverged from green algae over one billion years ago, and have various traits distinct from those of both green algae and land plants. Although most red algae are marine species (both unicellular and macrophytic), the Cyanidiales class of red algae includes unicellular species which live in hot springs, such as , which is a model species for biochemical and molecular biological studies. in red algae has previously been studied in intact cells. Here, we present the results of radiolabeling and stable isotope labeling experiments in intact plastids isolated from the unicellular red alga . We focused on two uncommon features: First, the galactose moiety of monogalactosyldiacylglycerol was efficiently labeled with bicarbonate, indicating that an unknown pathway for providing UDP-galactose exists within the plastid. Second, saturated fatty acids, namely, and stearic acids, were the sole products of fatty synthesis in the plastid, and they were efficiently exported. This finding suggests that the endoplasmic reticulum is the sole site of desaturation. We present a general principle of red algal biosynthesis, namely, \'indigenous C18 fatty acids are neither desaturated nor directly utilized within the plastid\'. We believe that this is valid in both lacking polyunsaturated fatty acids and marine red algae with a high content of arachidonic and eicosapentaenoic acids.

Keyword: fat metabolism

Milk production and nutrient digestibility responses to triglyceride or fatty supplements enriched in .

The objective of our study was to evaluate the effects of feeding triglyceride and fatty (FA) supplements enriched in (PA; C16:0) on production and nutrient digestibility responses of mid-lactation dairy cows. Fifteen Holstein cows (137 ± 49 d in milk) were randomly assigned to a treatment sequence in a 3 × 3 Latin square design. Treatments consisted of a control diet (CON; no added PA) or 1.5% FA added as either a FA supplement (PA-FA) or a triglyceride supplement (PA-TG). The PA supplements replaced soyhulls, and diets were balanced for glycerol content. Periods were 21 d in length with sample and data collection occurring during the final 5 d. Compared with CON, PA treatments increased dry matter (66.5 vs. 63.9%) and neutral detergent fiber (NDF) apparent digestibility (42.0 vs. 38.2%). Although PA treatments tended to increase 18-carbon FA apparent digestibility (79.1 vs. 77.9%), PA treatments decreased 16-carbon (63.1 vs. 75.8%) and total FA (72.0 vs. 76.5%) apparent digestibilities compared with CON. The PA treatments increased milk content (3.60 vs. 3.41%), milk yield (1.70 vs. 1.60 kg/d), yield of 16-carbon milk FA (570 vs. 471 g/d), 3.5% -corrected milk (47.6 vs. 46.5 kg/d), and energy-corrected milk (47.4 vs. 46.6 kg/d) compared with CON. The PA treatments did not affect dry matter intake (28.5 vs. 29.2 kg/d), milk yield (47.0 vs. 47.4 kg/d), milk protein yield (1.42 vs. 1.45 kg/d), milk lactose yield (2.29 vs. 2.31 kg/d), yield of <16-carbon milk FA (360 vs. 370 g/d), yield of >16-carbon milk FA (642 vs. 630 g/d), body weight (720 vs. 723 kg), or body condition score (3.14 vs. 3.23). We did not observe differences in digestibilities of dry matter, NDF, and 18-carbon FA between PA-TG and PA-FA. In contrast, PA-FA increased 16-carbon (68.6 vs. 57.6%) and total FA apparent digestibility (73.8 vs. 70.1%) compared with PA-TG. This resulted in PA-FA supplementation increasing the apparent digestibility of the PA supplement by ∼10 percentage points compared with PA-TG. Compared with PA-TG, PA-FA increased 16-carbon FA intake by 60 g/d, absorbed 16-carbon FA by 86 g/d, and absorbed total FA by 85 g/d. Compared with PA-TG, PA-FA increased dry matter intake (29.1 vs. 27.8 kg/d), yield of 16-carbon milk FA (596 vs. 545 g/d), and tended to increase milk yield (47.6 vs. 46.4 kg/d), milk yield (1.70 vs. 1.66 kg/d), and 3.5% -corrected milk (48.1 vs. 47.2 kg/d). In conclusion, the production response of dairy cows to PA tended to be greater for a FA supplement compared with a triglyceride supplement. Overall, PA increased NDF digestibility, milk yield, energy-corrected milk, and feed efficiency in mid-lactation dairy cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30\u202fmg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary as the same responses were induced by in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose in the obese mice.

Keyword: fat metabolism

Mp1 Protein, a Novel Virulence Factor, Carries Two Arachidonic -Binding Domains To Suppress Inflammatory Responses in Hosts.

infection causes talaromycosis (previously known as penicilliosis), a very important opportunistic systematic mycosis in immunocompromised patients. Different virulence mechanisms in have been proposed and investigated. In the sera of patients with talaromycosis, Mp1 protein (Mp1p), a secretory galactomannoprotein antigen with two tandem ligand-binding domains (Mp1p-LBD1 and Mp1p-LBD2), was found to be abundant. Mp1p-LBD2 was reported to possess a hydrophobic cavity to bind copurified (PLM). It was hypothesized that capturing of lipids from human hosts by expressing a large quantity of Mp1p is a virulence mechanism of It was shown that expression of Mp1p enhanced the intracellular survival of by suppressing proinflammatory responses. Mechanistic study of Mp1p-LBD2 suggested that arachidonic (AA), a precursor of paracrine signaling molecules for regulation of inflammatory responses, is the major physiological target of Mp1p-LBD2. In this study, we use crystallographic and biochemical techniques to further demonstrate that Mp1p-LBD1, the previously unsolved first binding domain of Mp1p, is also a strong AA-binding domain in Mp1p. These studies on Mp1p-LBD1 support the idea that the highly expressed Mp1p is an effective AA-capturing protein. Each Mp1p can bind up to 4 AA molecules. The crystal structure of Mp1p-LBD1-LBD2 has also been solved, showing that both LBDs are likely to function independently with a flexible linker between them. and potentially other pathogens highly expressing and secreting proteins similar to Mp1p can severely disturb host signaling cascades during proinflammatory responses by reducing the availabilities of important paracrine signaling molecules.Copyright © 2019 American Society for Microbiology.

Keyword: fat metabolism

Gibberellin Promotes Cell Growth and Induces Changes in Fatty Biosynthesis and Upregulates Fatty Biosynthetic Genes in Chlorella vulgaris UMT-M1.

Microalgae lipids and oils are potential candidates for renewable biofuels and nutritional inventions. Recent studies from our lab have shown that two plant hormones, auxin and jasmonic , influence microalgae growth and fatty accumulation. Therefore, in this study, a high oil-producing strain Chlorella vulgaris UMT-M1 was selected for hormonal study using gibberellin (GA). Exogenous GA was applied to early stationary culture of C. vulgaris UMT-M1. Results showed that GA gradually increases the cell density of C. vulgaris to up to 42% on days after treatment (DAT)-8 and also capable of delaying the algal senescence. However, the increment in cell density did not enhance the total oil production albeit transient modification of fatty compositions was observed for saturated (SFA) and polyunsaturated (PUFA) fatty acids. This illustrates that GA only promotes cell division and growth but not the oil accumulation. In addition, application of GA in culture medium was shown to promote transient increment of (C16:0) and stearic (C18:0) acids from DAT-4 to DAT-6 and these changes are correlated with the expression of β-ketoacyl ACP synthase I (KAS I) gene.

Keyword: fat metabolism

[The involvement of NOX1/NADPH oxidase in the development of non-alcoholic steatohepatitis].

Reactive oxygen species (ROS) are known to play a critical role in the development of non-alcoholic steatohepatitis (NASH). To clarify the source of ROS, we examined the expression of superoxide-generating NADPH oxidase isoforms in the liver of high- and high-cholesterol (HFC) diet-fed mice. The mRNA expression of NOX1 was significantly elevated in mice on HFC diet for 8 weeks. Increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 in HFC diet-fed wild-type mice (WT) were significantly ameliorated in mice deficient in Nox1 (Nox1-KO). Increased nitrotyrosine adduct formation, a marker of peroxynitrite-induced injury, was observed in hepatic sinusoids of WT, which was significantly suppressed in NOX1-KO. NOX1 mRNA was mainly expressed in liver sinusoidal endothelial cells (LSECs), and it was significantly up-regulated in primary cultured LSECs treated with (PA). The production of nitric oxide by LSECs and LSECs-dependent relaxation of hepatic stellate cells were significantly attenuated by PA treatment. In contrast, these effects of PA were not observed in cells isolated from Nox1-KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impair hepatic microcirculation through reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the development of NASH.

Keyword: fat metabolism

Dietary saturated fatty type impacts obesity-induced metabolic dysfunction and plasma lipidomic signatures in mice.

Saturated fatty (SFA) intake is associated with obesity, insulin resistance, and hepatic steatosis, but scant work examines the impact of SFA type upon these outcomes. We tested the hypothesis that an obesogenic diet prepared with medium chain SFA (MCSFA), mostly as lauric -derived from coconut oil, reduces obesity-induced outcomes compared to obesogenic diets prepared with increasing amounts long chain SFA (LCSFA), primarily . Mice were fed (16 weeks) a control, low diet or obesogenic diets prepared with differing content of MCSFA or LCSFA in which polyunsaturated and monounsaturated fatty acids (PUFA; MUFA) were kept constant. Inclusion of MCSFA in an obesogenic diet prevented hepatic accumulation and lowered indices of insulin resistance. Obesogenic diets reduced hepatic levels of de novo lipogenesis proteins (SCD1 and FASN) but elevated the adipose levels of mRNA for the pro-inflammatory markers Mcp-1 and Tnfα. Lipidomic analysis of plasma indicated that MCSFA intake resulted in a different lipidomic signature than LCSFA intake, prevented elevation of pro-inflammatory ceramides, but elevated concentrations of some lipids associated with elevated cardiovascular disease risk. Intake of the obesogenic diets in an SFA-type dependent manner elevated plasma concentrations of several phosphatidylcholine (PC) lipids having the long chain PUFA (LCPUFA) arachidonic (ARA) and docosahexaenoic (DHA), altered phospholipid ethers, and changed the triacylglyceryl environments of these LCPUFA. Our data indicate that (1) MCSFA reduce the severity of some obesogenic co-morbidities, (2) SFA-type modulates lipidomic signatures associated with cardiovascular disease and diabetes, and (3) dietary SFA type impacts LCPUFA .Published by Elsevier Inc.

Keyword: fat metabolism

The Spatiotemporal Deposition of Lysophosphatidylcholine Within Starch Granules of Maize Endosperm and its Relationships to the Expression of Genes Involved in Endoplasmic Reticulum-Amyloplast Trafficking and Galactolipid Synthesis.

The presence of lipids within starch granules is specific to cereal endosperm starches. These starch lipids are composed of lysophospholipids, especially lysophosphatidylcholine (LysoPC) and free fatty acids that strongly impact the assembly and properties of cereal starches. However, the molecular mechanisms associated with this specific routing have never been investigated. In this study, matrix-assisted laser desorption ionization mass spectrometry imaging revealed decreasing gradients in starch LysoPC concentrations from the periphery to the center of developing maize endosperms. This spatiotemporal deposition of starch LysoPC was similar to that previously observed for endoplasmic reticulum (ER)-synthesized storage proteins, i.e. zeins, suggesting that LysoPC might originate in the ER, as already reported for chloroplasts. Furthermore, a decrease of the palmitate concentration of amyloplast galactolipids was observed during endosperm development, correlated with the preferential trapping of palmitoyl-LysoPC by starch carbohydrates, suggesting a link between LysoPC and galactolipid synthesis. Using microarray, the homologous genes of the Arabidopsis ER-chloroplast trafficking and galactolipid synthesis pathways were also expressed in maize endosperm. These strong similarities suggest that the encoded enzymes and transporters are adapted to managing the differences between chloroplast and amyloplast homeostasis. Altogether, our results led us to propose a model where ER-amyloplast trafficking directs the LysoPC towards one of two routes, the first towards the stroma and starch granules and the other towards galactolipid synthesis.

Keyword: fat metabolism

Hepatocyte-Derived Lipotoxic Extracellular Vesicle Sphingosine 1-Phosphate Induces Macrophage Chemotaxis.

The pathophysiology of non-alcoholic steatohepatitis involves hepatocyte lipotoxicity due to excess saturated free fatty acids and concomitant proinflammatory macrophage effector responses. These include the infiltration of macrophages into hepatic cords in response to incompletely understood stimuli. Stressed hepatocytes release an increased number of extracellular vesicles (EVs), which are known to participate in intercellular signaling and coordination of the behavior of immune cell populations via their cargo. We hypothesized that hepatocyte-derived lipotoxic EVs that are enriched in sphingosine 1-phosphate (S1P) are effectors of macrophage infiltration in the hepatic microenvironment. Lipotoxic EVs were isolated from palmitate treated immortalized mouse hepatocytes and characterized by nanoparticle tracking analysis. Lipotoxic EV sphingolipids were quantified using tandem mass spectrometry. Wildtype and S1P receptor knockout bone marrow-derived macrophages were exposed to lipotoxic EV gradients in a microfluidic gradient generator. Macrophage migration toward EV gradients was captured by time-lapse microscopy and analyzed to determine directional migration. Fluorescence-activated cell sorting along with quantitative PCR and immunohistochemistry were utilized to characterize the cell surface expression of S1P receptor on intrahepatic leukocytes and hepatic expression of S1P receptor, respectively. Palmitate treatment induced the release of EVs. These EVs were enriched in S1P. Palmitate-induced S1P enriched EVs were chemoattractive to macrophages. EV S1P enrichment depended on the activity of sphingosine kinases 1 and 2, such that, pharmacological inhibition of sphingosine kinases 1 and 2 resulted in a significant reduction in EV S1P cargo without affecting the number of EVs released. When exposed to EVs derived from cells treated with palmitate in the presence of a pharmacologic inhibitor of sphingosine kinases 1 and 2, macrophages displayed diminished chemotactic behavior. To determine receptor-ligand specificity, we tested the migration responses of macrophages genetically deleted in the S1P receptor toward lipotoxic EVs. S1P receptor knockout macrophages displayed a marked reduction in their chemotactic responses toward lipotoxic palmitate-induced EVs. Palmitate-induced lipotoxic EVs are enriched in S1P through sphingosine kinases 1 and 2. S1P-enriched EVs activate persistent and directional macrophage chemotaxis mediated by the S1P receptor, a potential signaling axis for macrophage infiltration during hepatic lipotoxicity, and a potential therapeutic target for non-alcoholic steatohepatitis.

Keyword: fat metabolism

is a novel hypothalamic gene upregulated by a high- diet and leptin in mice.

Energy homeostasis is regulated by the hypothalamus but fails when animals are fed a high- diet (HFD), and leptin insensitivity and obesity develops. To elucidate the possible mechanisms underlying these effects, a microarray-based transcriptomics approach was used to identify novel genes regulated by HFD and leptin in the mouse hypothalamus.Mouse global array data identified as a novel gene highly upregulated by both a HFD and leptin challenge. In situ hybridisation showed expression upregulation by HFD and leptin in all major hypothalamic nuclei in agreement with transcriptomic gene expression data. Immunohistochemistry and studies in the hypothalamic clonal neuronal cell line, mHypoE-N42 (N42), confirmed that alpha 1-antichymotrypsin (αAC), the protein encoded by , is localised to neurons and revealed that it is secreted into the media. expression in N42 neurons is upregulated by and by leptin, together with and , and all three genes are downregulated by the anti-inflammatory monounsaturated , oleic . Additionally, palmitate upregulation of in N42 neurons is blocked by the NFκB inhibitor, BAY11, and the upregulation of expression in the hypothalamus by HFD is blunted in IL-1 receptor 1 knockout ( ) mice.These data demonstrate that expression is implicated in nutritionally mediated hypothalamic inflammation.

Keyword: fat metabolism

Berberine (BBR) Attenuated (PA)-Induced Lipotoxicity in Human HK-2 Cells by Promoting Peroxisome Proliferator-Activated Receptor α (PPAR-α).

BACKGROUND Berberine (BBR), a natural alkaloid isolated from Coptis chinensis, has frequently been reported as an antidiabetic reagent, partly due to its -lowering activity. Evidence suggests that BBR ameliorates palmitate-induced deposition and apoptosis in renal tubular epithelial cells (TECs), which tracks in tandem with the enhancement of peroxisome proliferator-activated receptor alpha (PPAR-alpha). The study aim was to investigate the roles of BBR in renal lipotoxicity in vitro, and investigate whether PPAR-alpha was the underlying mechanism. MATERIAL AND METHODS Human TECs (HK-2 cells) were injured with (PA), and then treated with BBR, BBR+PPAR-alpha inhibitor (GW6471), and PA+PPAR-alpha agonist (fenofibrate). Endoplasmic reticulum (ER) stress was assessed by measuring the expression of prospective evaluation of radial keratotomy (PERK), C/EBP-homologous protein (CHOP), and 78 kDa glucose-regulated protein (GRP78). was assessed by determining anabolism-associated genes, including fatty synthase (FAS), acetyl-CoA carboxylase (ACC), and lipoprotein lipase (LPL), as well as catabolism-associated gene, including carnitine palmitoyl transferase 1 (CPT1). Inflammatory response of HK-2 cells was evaluated by measuring interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha. Cell apoptosis and protein levels of cleaved-caspase-3 were evaluated. RESULTS PA downregulated PPAR-alpha and induced server lipotoxicity in HK-2 cells by ER stress, increasing deposition, and elevating inflammatory response of HK-2 cells accompanied with inducting cell apoptosis and cleaved-caspase-3, which were obviously reversed by additional treatment of BBR or PPAR-alpha agonist. However, the protective effect of BBR in PA-induced lipotoxicity in HK-2 cells was significantly ameliorated by PPAR-alpha inhibitor. CONCLUSIONS BBR attenuated PA-induced lipotoxicity via the PPAR-alpha pathway.

Keyword: fat metabolism

Acanthoic modulates lipogenesis in nonalcoholic fatty liver disease via FXR/LXRs-dependent manner.

Acanthoic (AA) is a pimaradiene diterpene isolated from Acanthopanax koreanum Nakai (Araliaceae), with anti-inflammatory and hepatic-protective effects. The present study intended to reveal the effect and mechanism of AA on nonalcoholic fatty liver disease (NAFLD) associated with accumulation by activating Farnesoid X receptor (FXR) and liver X receptors (LXRs) signaling. C57BL/6 mice were received a modified Lieber-DeCarli diet with 71% high- (L-D) and treated with AA (20 and 40\u202fmg/kg) or equal volume of saline for 12 weeks. The regulation of AA on accumulation was also detected in pro-steatotic stimulated AML12\u202fcells with (PA). When L-D diet-fed mice were treated with AA, loss in body weight, liver index, and liver droplet were observed along with reduced triglyceride (TG) and serum transaminase. Furthermore, AA decreased sterol regulatory element binding protein 1 (SREBP-1) and target genes expression, regulated PPARα and PPARγ expressions, ameliorated hepatic fibrosis markers, enhanced hepatic FXR and LXR, and regulated AMPK-LKB1 and SIRT1 signaling pathway. Moreover, AA attenuated accumulation via FXR and LXR activation in steatotic AML-12\u202fcells, which was confirmed by guggulsterones (FXR antagonist) or GW3965 (LXR agonist). Activation of FXR and LXR signaling caused by AA might increase AMPK-SIRT1 signaling and then contribute to modulating accumulation and fatty synthesis, which suggested that activated FXR-LXR axis by AA represented an effective strategy for relieving NAFLD.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Advanced Liver Fibrosis Is Independently Associated with and Insulin Levels in Patients with Non-Alcoholic Fatty Liver Disease.

Changes in occur during the development and progression non-alcoholic fatty liver disease (NAFLD). However, the fatty (FA) profile in red blood cells (RBC) from patients with liver fibrosis remains unexplored. Thus, the goal of this study was to evaluate the fatty profile in RBC, dietary intake and insulin resistance indicators in patients with NAFLD, according to the degree of hepatic fibrosis. Using elastography, patients were classified with ( = 52) and without ( = 37) advanced liver fibrosis. The fatty profile in RBC was analyzed using gas chromatography and the intake was evaluated through a 24-h dietary recall. Subjects with advanced liver fibrosis had higher levels of , stearic and oleic and total monounsaturated fatty (MUFA) and insulin ( < 0.05), and lower levels of elongase very long chain fatty acids protein-6 and the delta-5-desaturase enzymatic activity ( < 0.05). These results suggest a lack of regulation of enzymes related to FA in patients with advanced fibrosis.

Keyword: fat metabolism

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH).For in vitro studies, (PA) and free fatty (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200\u2009mg/kg/day) for 6 weeks. Histological staining\'s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative liver protein and gene expression levels were determined by Western Blot and qPCR, respectively.After establishing NASH models, LH treatment improved accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level.Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fat metabolism

pH effects on the and fatty acids accumulation in Chlamydomonas reinhardtii.

pH variations influence the delivery of essential nutrients and CO solubility, which impact algae . In this study the microalgal growth and chlorophyll, , and fatty acids content; along with the expression of some genes implicated in the biosynthesis of lipids were examined in Chlamydomonas reinhardtii subjected to pH values of 7.0, 7.8, and 8.5. At pH 7.8 an increase in cell growth was observed with a significant accumulation of chlorophyll (1.75-fold) when compared with growth at pH 7, while at pH 8.5 a sharp decrease in both parameters was observed when compared with the other pH values tested. content increased 3.0 (14.81% of dry cell weight, dcw) and 2.3 times (11.43% dcw) at pH 7.8 and 8.5, respectively, when compared with the experiment at pH 7 (4.97% dcw). The compositions of major fatty acids in the strains growing at pH 7.0, 7.8, or 8.5 were 25.7, 28.0, and 32.1% for ; 17.3, 14.7, and 25.7% for oleic ; and 9.8, 12.1, and 4.6% for linoleic ; respectively. qRT-PCR analysis showed that the transcripts of ß-carboxyltransferase, Acyl carrier protein 1, acyl-ACP thiolase 1, acyl-sn-glycerol-3-phosphate acyltransferase, and diacylglycerol acyl transferase isoform 3 were significantly induced at pH 7.8 when compared with the other two pH conditions. These results indicate that the induction of genes implicated in the early and final steps of biosynthesis contributes to their accumulation in the stationary phase. Our research suggests that a pH of 7.8 might be ideal to maximize growth and accumulation.© 2019 American Institute of Chemical Engineers.

Keyword: fat metabolism

d- chiro-Inositol Ameliorates High Diet-Induced Hepatic Steatosis and Insulin Resistance via PKCε-PI3K/AKT Pathway.

d- chiro-Inositol (DCI) is a biologically active component found in tartary buckwheat, which can reduce hyperglycemia and ameliorate insulin resistance. However, the mechanism underlying the antidiabetic effects of DCI remains largely unclear. This study investigated the effects and underlying molecular mechanisms of DCI on hepatic gluconeogenesis in mice fed a high diet and saturated -treated hepatocytes. DCI attenuated free fatty uptake by the liver via trafficking inhibition, reduced diacylglycerol deposition, and hepatic PKCε translocation. Thus, DCI could improve insulin sensitivity by suppressing hepatic gluconeogenesis. Subsequent analyses revealed that DCI decreased hepatic glucose output and the expression levels of PEPCK and G6 Pase in insulin resistant mice through PKCε-IRS/PI3K/AKT signaling pathway. Likewise, such effects of DCI were confirmed in HepG2 cells with palmitate-induced insulin resistance. These findings indicate a novel pathway by which DCI prevents hepatic gluconeogenesis, reduces deposition, and ameliorates insulin resistance via regulation of PKCε-PI3K/AKT axis.

Keyword: fat metabolism

Fatty acids uptake and oxidation are increased in the liver of rats with adjuvant-induced arthritis.

Severe rheumatoid cachexia is associated with pronounced loss of muscle and mass in patients with advanced rheumatoid arthritis. This condition is associated with dyslipidemia and predisposition to cardiovascular diseases. Circulating levels of triglycerides (TG) and free fatty acids (FFA) have not yet been consistently defined in severe arthritis. Similarly, the of these lipids in the arthritic liver has not yet been clarified. Aiming at filling these gaps this study presents a characterization of the circulating profile and of the fatty acids uptake and in perfused livers of rats with adjuvant-induced arthritis. The levels of TG and total cholesterol were reduced in both serum (10-20%) and liver (20-35%) of arthritic rats. The levels of circulating FFA were 40% higher in arthritic rats, possibly in consequence of cytokine-induced adipose tissue lipolysis. Hepatic uptake and oxidation of and oleic acids was higher in arthritic livers. The phenomenon results possibly from a more oxidized state of the arthritic liver. Indeed, NADPH/NADP and NADH/NAD ratios were 30% lower in arthritic livers, which additionally presented higher activities of the citric cycle driven by both endogenous and exogenous FFA. The lower levels of circulating and hepatic TG possibly are caused by an increased oxidation associated to a reduced synthesis of fatty acids in arthritic livers. These results reveal that the hepatic in arthritic rats presents a strong catabolic tendency, a condition that should contribute to the marked cachexia described for arthritic rats and possibly for the severe rheumatoid arthritis.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Effect of Hyperbaric Oxygen Therapy on Fatty Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Insulin-Dependent Diabetes Mellitus Patients: A Pilot Study.

Metabolic changes in insulin-dependent diabetes mellitus (IDDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in IDDM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in IDDM patients.Our study included 24 adult IDDM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4\xa0atmosphere absolute for 1\xa0hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis.HBOT decreased blood levels of total cholesterol (p<0.05), triglycerides (p<0.05) and low-density lipoprotein (p<0.05). HBOT increased plasma levels of individual FAs: (p<0.05), palmitoleic (p<0.05), docosapentaenoic (p<0.05) and docosahexaenoic (p<0.01), and decreased levels of stearic (p<0.05), alpha linolenic (p<0.05) and linoleic (p<0.01). Expression of IGFBP-1 (p<0.01) was increased, whereas the level of insulin (p<0.001) was decreased in the serum after HBOT.Our results indicate that HBOT exerts beneficial effects in IDDM patients by improving the profile and altering FA composition.Copyright © 2019 Canadian Diabetes Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high- diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and liver. PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free fatty acids during the clamp with Intralipid infusion reduced PAHSAs\' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial metabolic effects.

Keyword: fat metabolism

GCN2 deficiency ameliorates cardiac dysfunction in diabetic mice by reducing lipotoxicity and oxidative stress.

Excessive myocardial accumulation is a major feature of diabetic cardiomyopathy (DCM). Although general control nonderepressible 2 (GCN2) has been identified as a sensor of amino availability, it also functions as an important regulator of hepatic . Our previous studies have reported that GCN2 promotes pressure overload or doxorubicin-induced cardiac dysfunction by increasing cardiomyocyte apoptosis and myocardial oxidative stress. However, the impact of GCN2 on the development of DCM remains unclear. In this study, we investigated the effect of GCN2 on DCM in type 1 and type 2 diabetes animal models. After streptozotocin (STZ) or high- diet (HFD) plus low-dose STZ treatments, GCN2 mice developed less cardiac dysfunction, hyperlipidemia, myocardial hypertrophy, fibrosis, accumulation, oxidative stress, inflammation and apoptosis compared with wild-type (WT) mice. In diabetic hearts, GCN2 deficiency attenuated the upregulation of peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ), the phosphorylation of eIF2α and the induction of activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), as well as the reduction of Bcl-2. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, high glucose or -induced cell death, oxidative and endoplasmic reticulum stress and accumulation in H9C2 cells. Collectively, our data provide evidence that GCN2 deficiency protects cardiac function by reducing accumulation, oxidative stress and cell death. Our findings suggest that strategies to inhibit GCN2 activity in the heart may be novel approaches for DCM therapy.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

In vivo kinetic study of materno-fetal fatty transfer in obese and normal weight pregnant women.

Placental structure and function can be modified as a result of maternal obesity affecting materno-fetal fatty acids (FA) transport. We report for the first time, in humans and in vivo, the kinetics of placental FA transfer in normo-weight and in normolipemic obese pregnant women using stable isotopes. The administration of different tracer FA with similar behaviour to the mother at different time points allows the collection of kinetic information on materno-fetal transfer of FA despite only one sample of placenta and cord can be collected per subject. Computational modelling showed a good fit to the data when considering all maternal plasma classes but not when based only on non-esterified FA. The novel approach using multiple tracer FA administration combined with computational modelling shows a consistent time course of placental tracer FA and predicted total FA accumulation.We analyse for the first time the in vivo materno-fetal kinetic transfer of fatty acids (FA) labelled with stable isotopes in control and obese (OB) pregnant women. Labelled FA with a similar (stearic : C-SA; : C-PA; oleic : C-OA) were orally administered at -4\xa0h, -8\xa0h and -12\xa0h, respectively prior to elective caesarean section to 10 pregnant women with a body mass index >30 (OB) and 10 with a body mass index in the range 20-25 (NW). Placenta, venous and arterial cord blood were collected obtaining a wide range of FA enrichments. A combined experimental and computational modelling analysis was applied. FA fractional synthesis rate (FSR) in placenta was 11-12%\xa0h . No differences were observed between NW and normo-lipidemic OB. It was not possible to estimate FA FSR in cord blood with this oral bolus dose approach. Computational modelling demonstrated a good fit to the data when all maternal plasma classes were included but not with modelling based only on the non-esterified FA fraction. The estimated materno-fetal C-FA transfer was ∼1%. In conclusion, our approach using multiple C-FA tracers allowed us to estimated FSR in placental/maternal plasma but not in fetal/maternal compartments. Computational modelling showed a consistent time course of placental C-FA transfer and predicted total fetal FA accumulation during the experiment. We conclude that, in addition to non-esterified FA fraction in the maternal circulation, maternal plasma very low-density lipoprotein and other lipoproteins are important contributors to placental FA transfer to the fetus.© 2019 The Authors. The Journal of Physiology © 2019 The Physiological Society.

Keyword: fat metabolism

Wolf Extract Ameliorates Hepatic Steatosis through Regulation of , Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation.

Wolf (PCW) is an edible, pharmaceutical mushroom with remarkable biological properties including anti-tumor, anti-inflammation, anti-oxidation, anti-ageing, and anti-diabetic effects. In the current study, we investigated the effects of PCW extract on hepatic steatosis under in vitro and in vivo conditions, and elucidated the underlying mechanisms. In this study, a mixture of HepG2 cells treated with free fatty (FFA)- and oleic -and high- diet (HFD)-fed obese mice were used; in this background, the triglyceride (TG) levels in HepG2 cells and mice liver were measured, and the expression levels of genes associated with lipogenesis, fatty oxidation, endoplasmic reticulum (ER) stress, and autophagy were determined. Treatment of HepG2 cells with FFA enhanced intracellular TG levels in HepG2 cells, but co-treatment with PCW significantly attenuated the TG levels. Notably, PCW significantly enhanced the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein-1c (SREBP-1c) in FFA-treated HepG2 cells. PCW downregulated the expression of lipogenesis-related genes, but upregulated the expression of genes associated with fatty oxidation. Further, PCW inhibited FFA-induced expression of ER stress markers and induced autophagy proteins. However, inhibition of AMPK significantly attenuated the beneficial effects of PCW in HepG2 cells. Moreover, PCW efficiently decreased HFD-induced hepatic TG accumulation in vivo and increased the phosphorylation of hepatic AMPK. Three compounds present in PCW including poricoic , pachymic , and ergosterol, significantly decreased FFA-induced increase in intracellular TG levels, consistent with increased AMPK phosphorylation, suggesting that poricoic , pachymic , and ergosterol are responsible for PCW-mediated amelioration of hepatic steatosis. Taken together, these results demonstrated that PCW ameliorates hepatic steatosis through the regulation of , inhibition of ER stress, and activation of autophagy in an AMPK-dependent manner. This suggested that PCW can be potentially used for the treatment of hepatic steatosis.

Keyword: fat metabolism

Unsaturated Fatty Acids Increase the Expression of Hepassocin through a Signal Transducer and Activator of Transcription 3-Dependent Pathway in HepG2 Cells.

Hepassocin (HPS) is a hepatokine that regulates hepatocyte proliferation. It is known that HPS plays an important role in the development of nonalcoholic fatty liver diseases (NAFLD). Fatty acids, such as oleic (OLA), exhibit the ability to activate the signal transducer and activator of transcription-3 (STAT3), and the binding site of STAT3 is found in the promoter region of HPS. However, the regulation of HPS by fatty acids is still obscure. To clarify the regulation of HPS, we detected the expression of HPS by western blots. In addition, a hepatic steatosis cell culture model was established by treatment of different fatty acids, including linoleic (LNA), oleic , , and stearic . The intracellular accumulation was confirmed by oil red O staining. Blocking of STAT3 activity was achieved by the pretreatment of the STAT3 inhibitor, stattic. We found that activation of STAT3 by interleukin-6 (IL-6) was mediated in the regulation of HPS expression. Treatment of unsaturated fatty acids significantly induced intracellular accumulation in HepG2 cells. Moreover, the expressions of HPS were increased in unsaturated fatty -treated HepG2 cells, as compared with saturated fatty -treated groups. Also, the expression of HPS induced by OLA was blocked by the inhibition of STAT3 activity. Furthermore, we found that deletion of HPS by small interfering ribonucleic transfection decreased the protective effect of OLA on cell viability. Taken together, we provided evidence that STAT3 plays an important role in the regulation of OLA-induced HPS expression and the increased HPS may further participate in the development of NAFLD. In addition, the increase of HPS might be involved in the protective effect of OLA on cell viability.© 2018 AOCS.

Keyword: fat metabolism

Evidence for an alternative fatty desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated that enables them to synthesize, elongate and desaturate fatty acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target fatty and, in particular, fatty desaturation. This suggests that many cancer cells contain an unexplored plasticity in their fatty . Here we show that some cancer cells can exploit an alternative fatty desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human liver and lung carcinomas that desaturate palmitate to the unusual fatty sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known fatty desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in fatty desaturation and constitutes an unexplored metabolic rewiring in cancers.

Keyword: fat metabolism

Role of the mTOR‑FOXO1 pathway in obesity‑associated renal tubulointerstitial inflammation.

Since obesity is largely responsible for the growing incidence of renal tubulointerstitial inflammation, exploration into the mechanisms of obesity‑associated tubulointerstitial inflammation is essential. Studies have demonstrated that mammalian target of rapamycin\xa0(mTOR) is a crucial molecule in the pathogenesis of renal inflammation, including regulating the expression of inflammatory factors. The purpose of the present study was to further elucidate the role of mTOR in obesity‑associated tubulointerstitial inflammation. In the clinical study, obese and healthy subjects were recruited for physical examination, as well as the collection of blood and urine samples. Further study was performed on a high diet\xa0(HFD)‑induced obese rat model and a cultured human renal tubular epithelial cell line\xa0(HK‑2). The clinical study demonstrated that the participants with obesity had increased serum lipids, creatinine\xa0(Cr), urinary albumin to creatinine ratio\xa0(UACR) and urinary neutrophil gelatinase‑associated lipocalin\xa0(u‑NGAL). Moreover, the level of urinary monocyte chemoattractant protein‑1\xa0(u‑MCP‑1) was increased in the participants with obesity, and it was positively correlated with free fatty \xa0(FFA), UACR and u‑NGAL. In the in\xa0vivo study, the results indicated that the levels of serum lipids, Cr and blood urea nitrogen\xa0(BUN), as well as 24\xa0h urine protein and u‑NGAL, were significantly increased in the HFD‑fed obese rats. In addition, the infiltration of CD68+ cells into the renal interstitial area and the release of interleukin‑1β\xa0(IL‑1β) was observed in the kidneys of obese rats. Meanwhile, the supernatant from HK‑2 cells treated with stimulated THP‑1 monocyte migration. The upregulation of MCP‑1, phosphorylated forkhead boxO1 (p‑FOXO1), and phosphorylated mTOR (p‑mTOR) was observed in\xa0vivo and in\xa0vitro. However, inhibition of mTOR was able to alleviate the above effects. Overall, these results demonstrated that activated mTOR induced FOXO1 phosphorylation, which mediates renal MCP‑1 release, causes tubulointerstitial inflammation and ultimately leads to pathological renal changes and dysfunction. However, inhibition of mTOR may play a renoprotective role during the progression of obesity‑associated tubulointerstitial inflammation.

Keyword: fat metabolism

Cellular toxicity of dietary trans fatty acids and its correlation with ceramide and diglyceride accumulation.

High fatty (FA) levels are deleterious to pancreatic β-cells, largely due to the accumulation of biosynthetic intermediates, such as ceramides and diglycerides, which induce ER stress and apoptosis. Toxicity of palmitate (16:0) and oleate (18:1 cis-Δ) has been widely investigated, while very little data is available on the cell damages caused by elaidate (18:1 trans-Δ) and vaccenate (18:1 trans-Δ), although the potential health effects of these dietary trans fatty acids (TFAs) received great publicity. We compared the effects of these four FAs on cell viability, apoptosis, ER stress, JNK phosphorylation and autophagy as well as on ceramide and diglyceride contents in RINm5F insulinoma cells. Similarly to oleate and unlike palmitate, TFAs reduced cell viability only at higher concentration, and they had mild effects on ER stress, apoptosis and autophagy. Palmitate increased ceramide and diglyceride levels far more than any of the unsaturated fatty acids; however, incorporation of TFAs in ceramides and diglycerides was strikingly more pronounced than that of oleate. This indicates a correlation between the accumulation of intermediates and the severity of cell damage. Our findings reveal important metabolic characteristics of TFAs that might underlie a long term toxicity and hence deserve further investigation.Copyright © 2018. Published by Elsevier Ltd.

Keyword: fat metabolism

Liraglutide protects non-alcoholic fatty liver disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high- diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 diabetes mellitus (T2DM), has also been reported to decrease fatty degeneration of the liver. The purpose of this study is to explore whether liraglutide improves high- diet-induced non-alcoholic fatty liver disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the liver. After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg body weight) for four weeks, the liver, liver/body weight, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high- diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the liver of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high- diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Dynamics of Individual Fatty Acids in Muscle Stores and Membranes of a Songbird and Its Functional and Ecological Importance.

Although tissue fatty (FA) composition has been linked to whole-animal performance (e.g., aerobic endurance, metabolic rate, postexercise recovery) in a wide range of animal taxa, we do not adequately understand the pace of changes in FA composition and its implications for the ecology of animals. Therefore, we used a C to C diet shift experiment and compound-specific δC analysis to estimate the turnover rates of FAs in the polar and neutral fractions of flight muscle lipids (corresponding to membranes and droplets) of exercised and sedentary zebra finches (Taeniopygia guttata). Turnover was fastest for linoleic (LA; 18:2n6) and (PA; 16:0), with 95% replacement times of 10.8-17.7 d in the polar fraction and 17.2-32.8 d in the neutral fraction, but was unexpectedly slow for the long-chain polyunsaturated FAs (LC-PUFAs) arachidonic (20:4n6) and docosahexaenoic (22:6n3) in the polar fraction, with 95% replacement in 64.9-136.5 d. Polar fraction LA and PA turnover was significantly faster in exercised birds (95% replacement in 8.5-13.3 d). Our results suggest that FA turnover in intramuscular droplets is related to FA tissue concentrations and that turnover does not change in response to exercise. In contrast, we found that muscle membrane FA turnover is likely driven by a combination of selective LC-PUFA retention and consumption of shorter-chain FAs in energy . The unexpectedly fast turnover of membrane-associated FAs in muscle suggests that songbirds during migration could substantially remodel their membranes within a single migration stopover, and this may have substantial implications for how the FA composition of diet affects energy of birds during migration.

Keyword: fat metabolism

Trimethylamine N-Oxide Aggravates Liver Steatosis through Modulation of Bile and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Fatty Liver Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut microbiota, is associated with nonalcoholic fatty liver disease (NAFLD) and could influence bile (BA) . However, whether TMAO aggravates liver steatosis by modulating BA and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA . Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs liver function and increases hepatic triglyceride accumulation and lipogenesis in mice fed with a high- diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced lipogenesis in -treated HepG2 cells.TMAO aggravates liver steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fat metabolism

Gamma-glutamyl carboxylated Gas6 mediates the beneficial effect of vitamin K on lowering hyperlipidemia via regulating the AMPK/SREBP1/PPARα signaling cascade of .

The present study for the first time aims to examine the hypothesis that circulating gamma-glutamyl carboxylated growth arrest specific protein 6 (Gla-Gas6) deficiency may be associated with hyperlipidemia and vitamin K (VK) supplementation may ameliorate the impaired homeostasis via activating Gas6 protein. Subjects with hyperlipidemia (n=22) and age-matched healthy controls (n=19) were included in this study. Results showed that plasma levels of Gla-Gas6 protein and VK were significantly lower in hyperlipidemic subjects compared to control. Moreover, Gla-Gas6 levels were significantly and positively correlated with VK (P=.034, r=0.452) and negatively with triglyceride (P=.022, r=-0.485) and total cholesterol (P=.043, r=-0.435) in hyperlipidemic subjects, which suggest that VK supplementation may have a positive effect in activating Gas6 protein and thereby reducing the aberrant plasma levels. Further studies with high- diet (HFD)-fed animal model of hyperlipidemia demonstrated that VK supplementation (5 μg/kg body weight, 8 weeks) reduced the plasma levels, stimulated both the plasma levels and the hepatic protein expression of Gla-Gas6 protein, and regulated the AMPK/SREBP1/PPARα signaling pathways of hepatic in HFD-fed mice. Moreover, by using (PA, 0.75 mM)-treated both control and GGCX knockdown hepatocytes, this study dissected the direct role of Gla-Gas6 in mediating the positive effect of VK on preventing the PA-induced impaired hepatic via regulating AMPK/SREBP1/PPARα pathways. Combining all, the present study demonstrated the beneficial effect of VK supplementation in preventing the impaired homeostasis via activating VK-dependent Gas6 protein.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

A novel recombinant peptide INSR-IgG4Fc (Yiminsu) restores insulin sensitivity in experimental insulin resistance models.

Type 2 diabetes mellitus (T2DM) is a chronic degenerative endocrine and metabolic disease with high mortality and morbidity, yet lacks effective therapeutics. We recently generated a novel fusion peptide INSR-IgG4Fc, Yiminsu (YMS), to facilitate the high-affinity binding and transportation of insulin. Thus, the aim of the present study was to determine whether the novel recombinant peptide, YMS, could contribute to restoring insulin sensitivity and glycaemic control in insulin resistance models and revealing its underlying mechanism. (PA)-treated LO2 cells and high diet (HFD)-fed mice were treated with YMS. Therapeutic effects of YMS were measured using Western blotting, ELISA, qPCR, Histology and transmission electron microscopy. We observed that YMS treatment effectively improved insulin signaling in PA-treated LO2 cells and HFD-fed mice. Notably, YMS could significantly reduce serum levels of glucose, triglycerides, fatty acids and cholesterol without affecting the serum insulin levels. Moreover, our data demonstrated that YMS could restore glucose and homeostasis via facilitating insulin transportation and reactivating PI3K/Akt signaling in both PA-treated cells and liver, gastrocnemius and brown of HFD-fed mice. Additionally, we noticed that the therapeutic effects of YMS was similar as rosiglitazone, a well-recognized insulin sensitizer. Our findings suggested that YMS is a potentially candidate for pharmacotherapy for metabolic disorders associated with insulin resistance, particularly in T2DM.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: fat metabolism

[Glucagon-like peptide-1 regulates in hepatocytes through Foxo1/3].

Glucagon-like peptide-1 (GLP-1) has been reported to be effective in the treatment of nonalcoholic fatty liver disease (NAFLD). However, the molecular mechanism of GLP-1 on NAFLD is remained unclear. The present study was to detect whether the effect of GLP-1 on triglyceride (TG) content in hepatocytes is dependent on Foxos. HepG2 cells were treated with /oleic for 24 h. The knockdown of Foxo1, Foxo3 was conducted through small interfering RNA (siRNA). Real time PCT (RT-PCR) was used to detect the changes of the SREBP1c and Aco genes in HepG2 cells after Foxo1/3 knockdown. As expected, /oleic increased TG concentration in HepG2 cells [(12.65 ± 1.32) μg/mg vs. (4.32±0.54) μg/mg, 0.05]. Addition of GLP-1 dose (10, 50, 100nmol/L) dependently lowered the TG content and reached plateau at 100 nmol/L of GLP-1 [TG(8.38±1.47) μg/mg]. The GLP-1 effect on TG remained after knocking down either Foxo1 [(9.09±1.34)μg/mg] or Foxo3 [(8.90±1.60) μg/mg] alone, but not when knocking down Foxo1 and Foxo3 (Foxo1/3) together [(14.66±1.77)μg/mg]. Moreover, knocking down Foxo1/3 also abolished GLP-1 effect on SREBP1c and Aco expression. GLP-1 can inhibit the synthesis of TG in hepatocytes depending on Foxo1 and Foxo3. Further studies are needed to explore the specific mechanisms.

Keyword: fat metabolism

Encapsulation of soybean meal with fats enriched in and stearic acids: effects on rumen-undegraded protein and in vitro intestinal digestibility.

coating of soybean meal (SBM) can reduce its protein degradability in the rumen, but the encapsulation of SBM with (PA) and stearic acids (SA) has not yet been investigated, despite both fatty acids are common energy sources in dairy cow diets. This study aimed to evaluate the effects of applying a novel method, using either 400 or 500\xa0g /kg (treatments FL40 and FL50, respectively), which was enriched in PA and SA at different ratios (100:0, 75:25, 50:50, 25:75 and 0:100), on physical and chemical characteristics, ruminal degradability, solubility and in vitro intestinal protein digestibility (IVIPD) of the obtained products. Encapsulation of SBM in resulted in greater mean particle size and lower bulk density and protein solubility than unprotected SBM (USBM). Treatment FL50 resulted in increased (p <\xa00.01) rumen-undegraded protein (RUP) compared to USBM. There were no differences in RUP of SBM when different PA: SA ratios were used. The mean RUP content of treatments FL40 and FL50 (306 and 349\xa0g/kg, respectively) was greater compared to USBM (262\xa0g/kg, p <\xa00.05), but lower than that for a standard heat-treated SBM (431\xa0g/kg). Values of IVIPD did not differ among SBM, heat-treated SBM and FL40 and FL50 samples, all being greater than 97.8%. In conclusion, encapsulation of SBM with fats enriched in PA and SA proved to be effective in reducing protein solubility and increasing RUP without depressing protein digestibility in the intestine. For validation of the method, in vivo research to investigate the effects of these products on the production of dairy cows is warranted.

Keyword: fat metabolism

Myo-inositol alters 13C-labeled fatty in human placental explants.

We postulate that myo-inositol, a proposed intervention for gestational-diabetes, affects transplacental supply to the fetus. We investigated the effect of myo-inositol on fatty- processing in human placental-explants from uncomplicated pregnancies. Explants were incubated with 13C-labeled , 13C-oleic- and 13C-docosahexaenoic- across a range of myo-inositol concentrations for 24 h and 48 h. The incorporation of labeled-fatty-acids into individual lipids was quantified by liquid-chromatography-mass-spectrometry. At 24 h, myo-inositol increased the amount of 13C- and 13C-oleic- labeled lipids (median fold-change relative to control=1). Significant effects were seen with 30 µM myo-inositol (physiological) for 13C--lysophosphatidylcholines (1.26) and 13C--phosphatidylethanolamines (1.17). At 48 h, myo-inositol addition increased 13C-oleic--lipids but decreased 13C- and 13C-docosahexaenoic- lipids. Significant effects were seen with 30 µM myo-inositol for 13C-oleic--phosphatidylcholines (1.25), 13C-oleic--phosphatidylethanolamines (1.37) and 13C-oleic--triacylglycerols (1.32) and with 100 µM myo-inositol for 13C-docosahexaenoic--triacylglycerols (0.78). Lipids labeled with the same 13C-fatty- showed similar responses when tested at the same time-point, suggesting myo-inositol alters upstream processes such as fatty- uptake or activation. Myo-inositol supplementation may alter placental physiology with unknown clinical consequences.

Keyword: fat metabolism

Polysaccharides Prevent -Evoked Apoptosis and Autophagy in Intestinal Porcine Epithelial Cell Line via Restoration of Mitochondrial Function and Regulation of MAPK and AMPK/Akt/mTOR Signaling Pathway.

polysaccharide (GLP) extracted from (Leyss. ex Fr.) Karst, a traditional Chinese medicine, is a biologically active substance reported to possess anti-oxidative, anti-apoptotic, and neurological protection. However, it is unknown whether GLP have any protective effect against high- constituents-induced epithelial cell injury. The aim of this study was to investigate the protection and molecular mechanism of GLP on injury induced by (PA) in the intestinal porcine epithelial cell line (IPEC-J2). First, we tested whether the treatment of GLP attenuate PA-induced IPEC-J2 cell death. GLP markedly blocked PA-caused cytotoxicity and apoptosis in IPEC-J2 cells. Moreover, GLP recovered the decreased mitochondrial function and inhibited activation of caspase-dependent apoptotic pathway. Interestingly, PA promoted cell apoptosis and autophagy through stimulation of phosphorylation of mitogen-activated protein kinases (MAPKs), AMP-activated protein kinase (AMPK), and inhibition of phosphorylation of Akt and mammalian target of rapamycin (mTOR), which was reversed by GLP. Taken together, this study revealed a protective effect of GLP against PA-evoked IPEC-J2 cell death through anti-apoptotic and anti-autophagic properties.

Keyword: fat metabolism

Sodium tanshinone IIA sulfonate ameliorates hepatic steatosis by inhibiting lipogenesis and inflammation.

Non-alcoholic fatty liver disease (NAFLD) is becoming an epidemic disease in adults and children worldwide. Importantly, there are currently no approved treatments available for NAFLD. This study aims to investigate the potential applications of sodium tanshinone IIA sulfonate (STS) on improving the NAFLD condition using both in vitro and in vivo approaches. The results showed that STS markedly inhibited accumulation in oleic (OA) and (PA) treated HepG2 and primary immortalized human hepatic (PIH) cells. STS suppressed lipogenesis by inhibiting expression of sterol regulatory element binding transcription factor 1 (SREBF1), fatty synthase (FASN) and stearoyl-CoA desaturase (SCD). In addition, STS reduced inflammation in cells treated with OA-PA, shown by decreased transcriptional levels of tumor necrosis factor (TNF), transforming growth factor beta 1 (TGFB1) and interleukin 1 beta (IL1B). Consistently, protective effects on hepatic steatosis in db/db mice were observed after STS administration, demonstrated by decreased accumulation in mouse hepatocytes. This protective effect might be associated with STS induced activation of sirtuin 1 (SIRT1)/protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1) pathways. Our findings suggest a potential therapeutic role for STS in the treatment of NAFLD.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fat metabolism

Enhanced Fatty Scavenging and Glycerophospholipid Accompany Melanocyte Neoplasia Progression in Zebrafish.

Alterations in in cancer cells impact cell structure, signaling, and energy , making a potential diagnostic marker and therapeutic target. In this study, we combined PET, desorption electrospray ionization-mass spectrometry (DESI-MS), nonimaging MS, and transcriptomic analyses to interrogate changes in in a transgenic zebrafish model of oncogenic RAS-driven melanocyte neoplasia progression. Exogenous fatty uptake was detected in melanoma tumor nodules by PET using the surrogate tracer 14(R,S)-18F-fluoro-6-thia-heptadecanoic ([18F]-FTHA), consistent with upregulation of genes associated with fatty uptake found through microarray analysis. DESI-MS imaging revealed that FTHA uptake in tumors was heterogeneous. Transcriptome and lipidome analyses further highlighted dysregulation of glycerophospholipid pathways in melanoma tumor nodules, including increased abundance of phosphatidyl ethanolamine and phosphatidyl choline species, corroborated by DESI-MS, which again revealed heterogeneous phospholipid composition in tumors. Overexpression of the gene encoding lipoprotein lipase (LPL), which was upregulated in zebrafish melanocyte tumor nodules and expressed in the majority of human melanomas, accelerated progression of oncogenic RAS-driven melanocyte neoplasia in zebrafish. Depletion or antagonism of LPL suppressed human melanoma cell growth; this required simultaneous fatty synthase (FASN) inhibition when FASN expression was also elevated. Collectively, our findings implicate fatty acquisition as a possible therapeutic target in melanoma, and the methods we developed for monitoring fatty uptake have potential for diagnosis, patient stratification, and monitoring pharmacologic response. SIGNIFICANCE: These findings demonstrate the translational potential of monitoring fatty uptake and identify lipoprotein lipase as a potential therapeutic target in melanoma.©2019 American Association for Cancer Research.

Keyword: fat metabolism

Resveratrol Maintains Homeostasis via One of the Mechanisms Associated with the Key Circadian Regulator Bmal1.

Resveratrol (RES) possesses anti-inflammatory and anti-oxidant activities, and it can prevent liver disorders in obese and diabetic individuals. This study elucidated the mechanisms of brain and muscle Arnt-like protein-1 (Bmal1) in the protective effects of RES against liver disorders. The results indicated that RES ameliorated free fatty (FFA)-induced (oleic (OA): (PA) = 2:1) glycolipid metabolic disorders in hepatocytes. Simultaneously, RES partially reverted the relatively shallow daily oscillations of FFA-induced circadian clock gene transcription and protein expression in HepG2 cells. RES also attenuated FFA-triggered reactive oxygen species (ROS) secretion and restored mitochondrial membrane potential consumption, as well as the restoration of mitochondrial respiratory complex expression. This study provides compelling evidence that RES controls intracellular metabolic imbalance in a Bmal1-dependent manner. Overall, RES may serve as a promising natural nutraceutical for the regulation of metabolic disorders relevant to the circadian clock.

Keyword: fat metabolism

Diosgenin ameliorates -induced accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is characterized by excessive hepatic accumulation. Many studies have suggested that overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on energy , but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic fatty liver. The effects of DSG on , glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, consumption and synthesis in LO2 cells was investigated.The results indicated that induced obvious accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and fatty synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing fatty β-oxidation and decreasing synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: fat metabolism

Vascular endothelial growth factor B inhibits accumulation in C2C12 myotubes incubated with fatty acids.

To investigate (1) the effect of vascular endothelial growth factor B (VEGFB) on accumulation and the alteration of fatty acids and fatty -related enzymes in C2C12 myotubes incubated with fatty acids and (2) the regulatory effect of VEGFB on skeletal muscle . Mouse C2C12 myotubes were incubated with oleic (OA) and (PA), and differentiated mature C2C12 myotubes were treated with VEGFB. Oil-red O staining, BODIPY staining and cell triglycerides (TG) content were examined. Total RNA was isolated, and real-time PCR analysis was performed. Treatment with 100\u2009μM OA and 50\u2009μM PA induced droplet accumulation and increased TG content (\u2009<\u2009.01), and 100\u2009ng/mL VEGFB reduced droplet accumulation and decreased TG content (\u2009<\u2009.01). Treatment with 100\u2009ng/mL VEGFB significantly induced the mRNA expression of fatty transport protein 1 (FATP1) (\u2009<\u2009.01) and FATP4 (\u2009<\u2009.01). Treatment with 100\u2009ng/mL VEGFB significantly induced the mRNA expression of adipose TG lipase and hormone-sensitive lipase (\u2009<\u2009.01) as well as carnitine palmitoyltransferase I (\u2009<\u2009.01), peroxisome proliferator-activated receptor-γ coactivator-1α (\u2009<\u2009.01), acyl-coa dehydrogenase very long chain (\u2009<\u2009.05), acyl-coa synthetase long-chain family member 1 (\u2009<\u2009.01), peroxisomal acyl-coenzyme A oxidase 1 (\u2009<\u2009.05), and mitochondrial uncoupling protein 3 (\u2009<\u2009.01). VEGFB enhanced FATP1and FATP4 expression, promoted C2C12 myotube fatty oxidation and TG decomposition, and inhibited C2C12 myotube fatty re-esterification, thus inhibiting accumulation in C2C12 myotubes incubated with fatty acids.

Keyword: fat metabolism

Mitochondrial Metabolic Reprogramming by CD36 Signaling Drives Macrophage Inflammatory Responses.

A hallmark of chronic inflammatory disorders is persistence of pro-inflammatory macrophages in diseased tissues. In atherosclerosis this is associated with dyslipidemia and oxidative stress, but mechanisms linking these phenomena to macrophage activation remain incompletely understood. To investigate mechanisms linking dyslipidemia, oxidative stress and macrophage activation through modulation of immunemetabolism, and to explore therapeutic potential targeting specific metabolic pathways. Using a combination of biochemical, immunological, and ex vivo cell metabolic studies, we report that CD36 mediates a mitochondrial metabolic switch from oxidative phosphorylation to superoxide production in response to its ligand, oxLDL. Mitochondrial-specific inhibition of superoxide inhibited oxLDL-induced NF-κB activation and inflammatory cytokine generation. RNAseq, flow cytometry, H-labeled uptake, lipidomic analysis, confocal and EM imaging, and functional energetics revealed that oxLDL upregulated effectors of long-chain fatty (FA) uptake and mitochondrial import, while downregulating FA oxidation and inhibiting ATP5A, an electron transport chain (ETC) component. The combined effect is long-chain FA accumulation, alteration of mitochondrial structure and function, repurposing of the ETC to superoxide production, and NF-κB activation. Apoe null mice challenged with high diet showed similar metabolic changes in circulating Ly6C monocytes and peritoneal macrophages, along with increased CD36 expression. Moreover, mitochondrial ROS was positively correlated with CD36 expression in aortic lesional macrophages. These findings reveal that oxLDL/CD36 signaling in macrophages links dys-regulated FA to oxidative stress from the mitochondria, which drives chronic inflammation. Thus, targeting to CD36 and its downstream effectors may serve as potential new strategies against chronic inflammatory diseases such as atherosclerosis.

Keyword: fat metabolism

Soluble epoxide hydrolase derived mediators are elevated in bronchoalveolar lavage fluid from patients with sarcoidosis: a cross-sectional study.

Sarcoidosis is a systemic inflammatory multi-organ disease almost always affecting the lungs. The etiology remains unknown, but the hallmark of sarcoidosis is formation of non-caseating epithelioid cells granulomas in involved organs. In Scandinavia, >\u200930% of sarcoidosis patients have Löfgren\'s syndrome (LS), an acute disease onset mostly indicating a favorable prognosis. The impact of dysregulation of mediators, which has been investigated in other inflammatory disorders, is still unknown.Using three different liquid chromatography coupled to tandem mass spectrometry targeted platforms (LC-MS/MS), we quantified a broad suite of mediators including eicosanoids, sphingolipids and endocannabinoids in bronchoalveolar lavage (BAL) fluid from pulmonary sarcoidosis patients (n\u2009=\u200941) and healthy controls (n\u2009=\u200916).A total of 47 mediators were consistently detected in BAL fluid of patients and controls. After false discovery rate adjustment, two products of the soluble epoxide hydrolase (sEH) enzyme, 11,12-dihydroxyeicosa-5,8,14-trienoic (11,12-DiHETrE, p\u2009=\u20094.4E-5, q\u2009=\u20091.2E-3, median fold change\u2009=\u20096.0) and its regioisomer 14,15-dihydroxyeicosa-5,8,11-trienoic (14,15-DiHETrE, p\u2009=\u20093.6E-3, q\u2009=\u20093.2E-2, median fold change\u2009=\u20091.8) increased in patients with sarcoidosis. Additional shifts were observed in sphingolipid , with a significant increase in -derived sphingomyelin (SM16:0, p\u2009=\u20091.3E-3, q\u2009=\u20091.7E-2, median fold change\u2009=\u20091.3). No associations were found between these 3 mediators and LS, whereas levels of SM 16:0 and 11,12-DiHETrE associated with radiological stage (p\u2009<\u20090.05), and levels of 14,15-DiHETrE were associated with the BAL fluid CD4/CD8 ratio.These observed shifts in mediators provide new insights into the pathobiology of sarcoidosis and in particular highlight the sEH pathway to be dysregulated in disease.

Keyword: fat metabolism

Energy and nitrogen partitioning in dairy cows at low or high metabolizable protein levels is affected differently by postrumen glucogenic and lipogenic substrates.

This study tested the effects of energy from glucogenic (glucose; GG) or lipogenic (palm olein; LG) substrates at low (LMP) and high (HMP) metabolizable protein levels on whole-body energy and N partitioning of dairy cattle. Six rumen-fistulated, second-lactation Holstein-Friesian dairy cows (97 ± 13 d in milk) were randomly assigned to a 6 × 6 Latin square design in which each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of rest. A total mixed ration consisting of 42% corn silage, 31% grass silage, and 27% concentrate (dry matter basis) was formulated to meet 100 and 83% of net energy and metabolizable protein requirements, respectively, and was fed at 90% of ad libitum intake by individual cow. Abomasal infusion treatments were saline (LMP-C), isoenergetic infusions (digestible energy basis) of 1,319 g/d of glucose (LMP-GG), 676 g/d of palm olein (LMP-LG; major fatty constituents are , oleic, and linoleic ), or 844 g/d of essential AA (HMP-C), or isoenergetic infusions of 1,319 g/d of glucose + 844 g/d of essential AA (HMP-GG) or 676 g/d of palm olein + 844 g/d of essential AA (HMP-LG). The experiment was conducted in climate respiration chambers to determine energy and N balance in conjunction with milk production and composition, nutrient digestibility, and plasma constituents. Infusion of GG and LG decreased dry matter intake, but total gross energy intake from the diet plus infusions was not affected by GG or LG. Furthermore, GG or LG did not affect total milk, protein, or lactose yields. Infusing GG or LG at the HMP level did not affect milk production differently than at the LMP level. Infusion of GG stimulated energy retention in body tissue, increased plasma glucose and insulin concentrations, decreased lipogenic metabolites in plasma, and decreased milk yield and milk energy output. Nitrogen intake decreased and milk N efficiency increased in response to GG, and N retention was not affected. Infusion of LG tended to increase metabolizable energy intake, increased milk yield and milk energy output, increased plasma triacylglycerides and long-chain fatty concentrations, and had no effect on energy retention. Infusion of LG decreased N intake but did not affect milk N efficiency or N retention. Compared with the LMP level, the HMP level increased dry matter intake, gross and metabolizable energy intake, and total milk, , protein, and lactose yields. Milk energy output increased at the HMP level, and protein level did not affect total energy retention. Heat production increased at the HMP level, but only when GG and LG were infused. The HMP level increased N intake, milk N output, and plasma urea concentration, tended to increase N retention, and decreased milk N efficiency. Regardless of protein level, GG promoted energy retention and improved milk N efficiency, but not through increased milk protein yield. Infusion of LG partitioned extra energy intake into milk and had no effect on milk N efficiency.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Apoptotic germ cells regulate Sertoli cell storage and fatty oxidation

The presence of droplets (LDs) and the utilization of fatty acids (FAs) as a source of energy are Sertoli cell (SC) putative characteristics. It is well known that SCs can phagocyte and degrade apoptotic germ cells (AGCs) resulting in increasing content and ATP levels. A relationship between the regulation of storage and of oxidation in SC might be envisaged. The aim of this study was to analyze whether AGCs and FAs are able to simultaneously regulate molecular mechanisms involved in storage and in FA oxidation in SC. The experimental model utilized in this study consisted in SC cultures obtained from 20-day-old rats that were co-cultured with AGC or treated with (PA, 500 μM) for 24 and 48 h. AGC and PA increase LD, triacylglycerol (TAG) content and mRNA levels of Plin1, Plin2, Plin3 (proteins involved in TAG storage). Simultaneously, AGC and PA rise the extent of FA oxidation and mRNA levels of Cpt1 and Lcad (proteins involved in FA degradation). Results also show that peroxisome proliferator-activated receptor (PPAR) transcriptional activity, transcription factor which participate in regulation, increases by AGC and PA treatment in SC. Additionally, the presence of a PPARg antagonist decreases the upregulation of LD content and Plin1 expression. Similarly, the presence of a PPARb/d antagonist reduces the increase in FA oxidation and Cpt1 mRNA levels. Altogether these results suggest that AGC and FA, which probably generate PPAR ligands, regulate storage and fatty utilization, contributing to the energy homeostasis in the seminiferous tubules.

Keyword: fat metabolism

Chronic intake of moderate -enriched diet induces fatty liver and low-grade inflammation without obesity in rabbits.

Non-Alcoholic Fatty Liver Disease (NAFLD) is the cause of chronic liver disease. Even though NAFLD is strongly associated with obesity and metabolic syndrome, there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate -enriched diet on hepatic accumulation and their relationship with inflammation. Rabbits fed with a moderate Fatty-- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and liver function. Liver samples were analyzed by histology and RT-qPCR to measure accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate metabolic changes, but it induced fatty liver. We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of -enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of metabolic disorders, such as overweight/obesity and metabolic syndrome. However, the excessive intake of fatty acids renders these mechanisms less efficient for delaying the start of metabolic alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: fat metabolism

Stable Isotope-Labeled Lipidomics to Unravel the Heterogeneous Development Lipotoxicity.

Non-alcoholic fatty liver disease (NAFLD) as a global health problem has clinical manifestations ranging from simple non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH), cirrhosis, and cancer. The role of different types of fatty acids in driving the early progression of NAFL to NASH is not understood. overload causing lipotoxicity and inflammation has been considered as an essential pathogenic factor. To correlate the profiles with cellular lipotoxicity, we utilized (C16:0)- and especially unprecedented palmitoleic (C16:1)-induced overload HepG2 cell models coupled with lipidomic technology involving labeling with stable isotopes. C16:0 induced inflammation and cell death, whereas C16:1 induced significant droplet accumulation. Moreover, inhibition of de novo sphingolipid synthesis by myriocin (Myr) aggravated C16:0 induced lipoapoptosis. profiles are different in C16:0 and C16:1-treated cells. Stable isotope-labeled lipidomics elucidates the roles of specific fatty acids that affect and cause lipotoxicity or droplet formation. It indicates that not only saturation or monounsaturation of fatty acids plays a role in hepatic lipotoxicity but also Myr inhibition exasperates lipoapoptosis through ceramide in-direct pathway. Using the techniques presented in this study, we can potentially investigate the mechanism of and the heterogeneous development of NAFLD.

Keyword: fat metabolism

production by Lipomyces starkeyi using sap squeezed from felled old oil palm trunks.

The ability of oleaginous yeast Lipomyces starkeyi to efficiently produce lipids when cultivated on sap extracted from felled oil palm trunk (OPT) as a novel inexpensive renewable carbon source was evaluated. OPT sap was found to contain approximately 98\xa0g/L glucose and 32\xa0g/L fructose. Batch fermentations were performed using three different OPT sap medium conditions: regular sap, enriched sap, and enriched sap at pH 5.0. Under all sap medium conditions, the cell biomass and production achieved were approximately 30\xa0g/L and 60% (w/w), respectively. L.\xa0starkeyi tolerated acidified medium (initial pH ≈ 3) and produced considerable amounts of ethanol as well as xylitol as by-products. The fatty profile of L.\xa0starkeyi was remarkably similar to that of palm oil, one of the most common vegetable oil feedstock used in biodiesel production with oleic as the major fatty followed by , stearic and linoleic acids.Copyright © 2019 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and Liver in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans fatty acids. The effect of interesterified fats containing or stearic acids on and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high- diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in and inflammatory processes in liver and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while liver collagen content was determined by Sirius Red staining. Both interesterified fats increased liver collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: fat metabolism

Fatty Profile and Cardiometabolic Markers in Relation with Diet Type and Omega-3 Supplementation in Spanish Vegetarians.

Plant-based diets are becoming increasingly popular, and scientific information concerning the nutritional status in this population is needed. This study determined the fatty profile of Spanish lacto-ovo vegetarians (LO-vegetarians) and vegans. Participants were 104 healthy adults, LO-vegetarians ( = 49) and vegans ( = 55). Lifestyle habits and consumption of food and omega-3 supplements were estimated by questionnaires. BMI, blood pressure, and abdominal and body were determined. Serum was collected to analyze fatty acids, glucose, lipids, homocysteine, insulin, and leptin. Volunteers were classified according to serum omega-6 to omega-3 (-6/-3) ratio into three groups: -6/-3 < 10, -6/-3 ≥ 10 to 20, and -6/-3 > 20. Results showed low cardiovascular risk and high insulin sensitivity with negligible differences between diet types. Linoleic (C18:2-6) was the major serum fatty , followed by oleic (C18:1-9) and (C16:0) acids. In contrast, serum eicosapentaenoic (EPA, C20:5-3) and docosahexaenoic (DHA, C22:6-3) were (median, interquartile range) 0.27, 0.18% and 1.59, and 0.93%, respectively. Users of -3 supplements (<10% of total vegetarians) had significantly higher EPA than non-users, while frequent consumption of flax-seeds was associated with increased α-linolenic (C18:3-3). However, neither -3 supplementation nor food consumption affected DHA levels in this vegetarian population.

Keyword: fat metabolism

Berberine attenuates sodium palmitate-induced accumulation, oxidative stress and apoptosis in grass carp(Ctenopharyngodon idella)hepatocyte in vitro.

The objective of this work was to investigate the effect of berberine (BBR) on the Cell viability, accumulation, apoptosis, cytochrome c, caspase-9 and caspase-3 in accumulation-hepatocytes induced by sodium palmitate in vitro. The accumulation-hepatocytes (induced by 0.5\u202fmM sodium palmitate for 24\u202fh) were treated with 5\u202fμM berberine for 12\u202fh. Then, the Cell viability, intracellular triglyceride (TG) content, peroxide (LPO), malonaldehyde (MDA) content, cytochrome c, caspase-9, caspase-3 and apoptosis were detected. Sodium palmitate decreased Cell viability and increased intracellular TG content, droplet accumulation, LPO and MDA concentrations, caused caspase-3 and caspase-9 activation, then led to apoptosis accompanied by cytochrome c release from mitochondria into the cytoplasm. Beberine could improve intracellular droplet accumulation and oxidative stress, while reduce apoptosis induced by sodium palmitate.Copyright © 2019. Published by Elsevier Ltd.

Keyword: fat metabolism

Attenuating effect of silibinin on -induced apoptosis and mitochondrial dysfunction in pancreatic β-cells is mediated by estrogen receptor alpha.

High levels of circulating free fatty acids often trigger pancreatic β cell dysfunction during the development of type 2 diabetes. Silibinin, the main component of Silybum marianum fruit extract (silymarin), is reported to have anti-diabetic effect. This study is designed to determine the protective effect of silibinin on -induced damage in a rat pancreatic β-cell line, INS-1 cells. Our results demonstrate that silibinin improves cell viability, enhances insulin synthesis and secretion, and resumes normal mitochondrial function in -treated INS-1 cells. An accumulating body of evidence has shown that the estrogen receptors are key molecules involved in glucose and . Our results suggest that silibinin upregulates ERα signaling pathway from the finding that ERα-specific inhibitors abolish the anti-lipotoxic effect of silibinin. In conclusion, these findings suggest that silibinin protects INS-1 cells against apoptosis and mitochondrial damage through upregulation of ERα pathway.

Keyword: fat metabolism

lncRNA MEG3 promotes hepatic insulin resistance by serving as a competing endogenous RNA of miR-214 to regulate ATF4 expression.

MicroRNA (miR)‑214 has been demonstrated to suppress gluconeogenesis by targeting activating transcription factor 4 (ATF4), which regulates gluconeogenesis by affecting the transcriptional activity of forkhead box protein O1 (FoxO1). Our previous study revealed that the upregulation of maternally expressed gene 3 (MEG3), a long noncoding RNA, enhanced hepatic insulin resistance via increased FoxO1 expression. The present study aimed to explore whether miR‑214 and ATF4 were involved in the MEG3‑mediated increase of FoxO1 expression. MEG3, miR‑214 and ATF4 expression were examined by reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among MEG3, miR‑214 and ATF4 was analysed using the luciferase reporter assay. MEG3‑targeting small interference RNAs were injected into high‑ diet (HFD)‑fed mice to verify the role of MEG3 in hepatic insulin resistance in\xa0vivo. MEG‑3 and ATF4 were demonstrated to be upregulated and miR‑214 was indicated to be downregulated in the livers of HFD‑fed and ob/ob mice. In mouse primary hepatocytes, palmitate time‑dependently increased MEG3 and ATF4 but decreased miR‑214 expression levels. Furthermore, MEG3 served as a competing endogenous RNA (ceRNA) for miR‑214 to facilitate ATF4 expression, while miR‑214 inhibition and ATF4 overexpression reversed the MEG3 knockdown‑mediated decrease in the expression of FoxO1 and FoxO1‑downstream targets phosphoenolpyruvate carboxykinase and glucose‑6‑phosphatase catalytic subunit. In HFD‑fed mice, MEG3 knockdown substantially improved impaired glucose and insulin tolerance, while downregulating HFD‑induced ATF4 expression and upregulating HFD‑suppressed miR‑214 expression. In conclusion, MEG3 promoted hepatic insulin resistance by serving as a ceRNA of miR‑214 to facilitate ATF4 expression. These data provide insight into the molecular mechanism of MEG3 involvement in the development of type 2 diabetes mellitus.

Keyword: fat metabolism

Effects of timing of supplementation during early lactation on nutrient digestibility, energy balance, and of dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation during early lactation on nutrient digestibility, energy intake and balance, and metabolic responses of dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh (FR) period (1-24 d in milk) cows were assigned to either a control diet containing no supplemental (CON) or a C16:0-supplemented diet [PA; 1.5% of diet dry matter (DM)]. During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA diet (1.5% of diet DM) in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, compared with CON, PA increased DM digestibility by 3.0 percentage units and neutral detergent fiber (NDF) digestibility by 4.4 percentage units, and the increase in these variables was consistent over time. Although PA did not affect 18-carbon fatty (FA) digestibility, it decreased 16-carbon FA digestibility by 10.8 percentage units and total FA digestibility by 4.7 percentage units compared with CON. We observed a tendency for an interaction between treatment and time for total FA digestibility and 16-carbon FA digestibility due to the difference in FA digestibility between PA and CON reducing over time. Compared with CON, PA increased digestible energy intake by 3.9 Mcal/d, metabolizable energy intake by 3.5 Mcal/d, and net energy for lactation intake by 2.5 Mcal/d. The PA diet also increased milk energy output, negative energy balance, and plasma nonesterified fatty concentration and reduced plasma insulin concentration. We also observed a tendency for an interaction between treatment and time for energy balance due to cows receiving the PA treatment being in a greater negative energy balance over time. During the PK period, PA increased DM digestibility by 2.9 percentage units and NDF digestibility by 3.5 percentage units compared with CON. Although PA decreased 16-carbon FA digestibility by 7.0 percentage units, PA did not affect 18-carbon FA digestibility or total FA digestibility. Feeding PA during the PK period increased energy intake and milk energy output and did not affect energy balance. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased DM and NDF digestibilities and energy intake compared with a control diet containing no supplemental . Feeding C16:0 markedly increased milk energy output in both the FR and PK periods but increased negative energy balance only in the FR period.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Glycerolipid Composition of the Red Macroalga and Comparison to the Closely Related Producing Different Types of Eicosanoids.

The red macroalga is a well-known producer of eicosanoids such as hydroxyeicosatetraenoic acids, but the alga produces almost no prostaglandins, unlike the closely related . This indicates that the related two algae would have different enzyme systems or substrate composition. To carry out more in-depth discussions on the metabolic pathway of eicosanoids between the two algae, we investigated the characteristics of glycerolipids, which are the substrates of eicosanoids production, of and compared them to the reported values of . In , monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol (SQDG), and phosphatidylcholine (PC) were the major classes and accounted for 44.4% of the total extract. The predominant fatty acids were arachidonic (20:4n-6), an eicosanoids precursor, and (16:0). The 20:4n-6 content was extremely high in MGDG and PC (>70%), and the 16:0 content was extremely high in DGDG and SQDG (>40%). A chiral-phase HPLC analysis showed that fatty acids were esterified at the -1 and -2 positions of those lipids. The glycerolipid molecular species were determined by reversed-phase HPLC⁻ESI⁻MS analysis. The main glycerolipid molecular species were 20:4n-6/20:4n-6 (-1/-2) for MGDG (63.8%) and PC (48.2%), 20:4n-6/16:0 for DGDG (71.1%) and SQDG (29.4%). These characteristics of were almost the same as those of . Hence, the differences of the eicosanoids producing ability between the two algae would not be due to the difference of substrate composition but the difference of enzyme system.

Keyword: fat metabolism

Electric pulse stimulation inhibited accumulation on C2C12 myotubes incubated with oleic and .

To investigate the effect of electrical pulse stimulation (EPS) on accumulation and alteration of fatty -related enzymes in C2C12 myotubes incubated with fatty acids. Mouse C2C12 myotubes were incubated with oleic and , and differentiated C2C12 myotubes were treated with EPS, oil-red O (ORO), BODIPY staining and triglyceride (TG) content were examined. Total RNA was isolated, and real-time polymerase chain reaction analysis was performed. (1) EPS decreased TG content (\u2009<\u2009.01). (2) EPS significantly induced the mRNA expression of FAD/CD36 (\u2009<\u2009.05), FATP4 (\u2009<\u2009.001), FABP1 (\u2009<\u2009.01) and FABP5 (\u2009<\u2009.01). (3) EPS significantly inhibited the mRNA expression of fatty synthase (\u2009<\u2009.01). (4) Adipose triglyceride lipase and hormone-sensitive lipase expression were significantly elevated (\u2009<\u2009.001), and induced the mRNA expression of CPT1 (\u2009<\u2009.01), ACOX1 (\u2009<\u2009.05), UCP3 (\u2009<\u2009.05) and PPARα (\u2009<\u2009.001) after EPS. EPS reduced droplet accumulation; enhanced CD36, FATP4, FABP1 and FABP5 expression; inhibited C2C12 myotube fatty re-esterification; and promoted fatty oxidation in C2C12 myotubes.

Keyword: fat metabolism

Optimization of Synechococcus sp. VDW Cultivation with Artificially Prepared Shrimp Wastewater for Ammonium Removal and Its Potential for Use As a Biofuel Feedstock.

To investigate the potential of application of marine cyanobacterium for concurrent biomass production and ammonium removal, Synechococcus sp. VDW was cultured under different conditions in medium containing varying concentrations of NHCl. Response surface methodology (RSM) was then used to build a predictive model of the combined effects of independent variables (pH, inoculum size, ammonium concentration). At the optimum conditions of initial pH 7.4, inoculum size 0.17 (OD730) and ammonium concentration 10.5 mg L, the maximum ammonium removal and biomass productivity were about 95% and 34 mg Ld, respectively, after seven days of cultivation. The result of fatty methyl ester (FAME) analysis showed that the major fatty acids were (C16:0), linoleic (C18:2 n6 cis), palmitoleic (C16:1) and oleic (C18:1 n9 cis), which accounted for more than 80% weight of total fatty acids. Further, analysis of neutral accumulation using flow cytometry revealed that the mean of the fluorescence intensity increased under optimal conditions. These results indicate that Synechococcus sp. VDW has the potential for use for concurrent water treatment and production of biomass that can be applied as biofuel feedstock.

Keyword: fat metabolism

Effects of timing of supplementation on production responses of early-lactation dairy cows.

The objective of our study was to evaluate the effects of timing of (C16:0) supplementation on production responses of early-lactation dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh period (FR; 1-24 d in milk) cows were assigned to either a control diet containing no supplemental (CON) or a diet supplemented with C16:0 (, PA; 1.5% of diet dry matter). During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA (1.5% of diet dry matter) diet in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, we did not observe treatment differences for dry matter intake or milk yield. Compared with CON, PA increased the yield of 3.5% -corrected milk by 5.30 kg/d, yield of energy-corrected milk (ECM) by 4.70 kg/d, milk content by 0.41% units, milk yield by 280 g/d, and protein yield by 100 g/d. The increase in milk associated with the PA treatment during the FR period occurred due to an increase in yield of 16-carbon milk fatty acids (FA) by 147 g/d (derived from both de novo synthesis and extraction from plasma) and preformed milk FA by 96 g/d. Compared with CON, PA decreased body weight (BW) by 21 kg and body condition score (BCS) by 0.09 units and tended to increase BW loss by 0.76 kg/d. Although PA consistently increased milk yield and ECM over time, a treatment × time interaction was observed for BW and BCS due to PA inducing a greater decrease in BW and BCS after the second week of treatments. Feeding PA during the PK period increased milk yield by 3.45 kg/d, yield of 3.5% -corrected milk by 4.50 kg/d, yield of ECM by 4.60 kg/d, milk content by 0.22% units, milk yield by 210 g/d, protein yield by 140 g/d, and lactose yield by 100 g/d but tended to reduce BW by 10 kg compared with CON. Also, during the PK period we observed an interaction between diet fed in the FR and PK periods for milk yield due to feeding PA during the PK period increasing milk yield to a greater extent in cows that received the CON diet (+240 g/d) rather than the PA diet (+180 g/d) during the FR period. This difference was associated with the yield of preformed FA because feeding PA during the PK period increased the yield of preformed milk FA only in cows that received the CON diet during the FR period. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased the yield of ECM in both the FR and PK periods compared with a control diet. For some variables, the effect of feeding C16:0 was affected by timing of supplementation because milk yield increased only during the PK period and BW decreased to a greater extent in the FR period. Regardless of diet fed in the FR period, feeding a C16:0 supplement during the PK period increased yields of milk and milk components.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Increased Dynamin-Related Protein 1-Dependent Mitochondrial Fission Contributes to High--Diet-Induced Cardiac Dysfunction and Insulin Resistance by Elevating Tafazzin in Mouse Hearts.

High (HF)-diet-induced insulin resistance is a major contributor to the pathogenesis of cardiovascular diseases. However, the molecular mechanisms that regulate cardiac insulin signaling are not fully understood. The regulatory role of tafazzin in the hearts of HF-diet-fed mice is investigated.Mice are fed a HF diet or low (LF) diet for up to 24\xa0weeks. After 24\xa0weeks, it is found that HF-diet-induced cardiac dysfunction is linked to overexpression of the mitochondrial protein tafazzin. Increased tafazzin promotes mitochondrial fission and impairs insulin signaling, which is mediated by dynamin-related protein 1 (Drp-1) translocation from the cytosol to the mitochondria. Furthermore, knockdown of tafazzin with siRNA inhibits -induced mitochondrial fission and restores insulin sensitivity. Moreover, miR-125b-5p as an upstream regulator targeting tafazzin is identified and palmitate-induced insulin resistance further rescued.In HF-diet-fed mouse hearts, increased tafazzin contributes to insulin resistance via mediating Drp-1 translocation to the mitochondria, and a small non-coding RNA, miR-125b-5p, at least partially regulates this signaling pathway and alleviates insulin resistance.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fat metabolism

Lycopus lucidus Turcz. ex Benth. Attenuates free fatty -induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high- diet-induced obese mice.

Non-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free fatty (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high- diet (HFD; 60 kcal% ) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative liver weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic .LLE significantly decreased the intracellular accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative liver weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: fat metabolism

Effects of Chlorpyrifos on Cholinesterase and Serine Lipase Activities and in Brains of Rainbow Trout (Oncorhynchus mykiss).

Chlorpyrifos is an organophosphorus insecticide that elicits acute toxicity through inhibition of acetylcholinesterase (AChE), leading to acetylcholine accumulation and prolonged stimulation of cholinergic receptors throughout the central and peripheral nervous systems. Previous studies have indicated that neurodevelopment may also be impaired through alternative pathways, including reduction of cAMP catalyzed downstream events. The upstream initiating events that underlie non-cholinergic neurological actions of chlorpyrifos and other organophosphorus compounds remain unclear. To investigate the potential role of disruption of fatty signaling as a mechanism of toxicity, and fatty profiles were examined to identify alterations that may play a critical role in upstream signaling in the CNS. Juvenile rainbow trout were treated for 7 days with nominal chlorpyrifos concentrations previously reported to diminish olfactory responses (10, 20, and 40\u2009μg/L). While lethality was noted higher doses, measured chlorpyrifos concentrations of 1.38\u2009μg/L (nominal concentration 10\u2009μg/L) significantly reduced the activity of AChE and two serine lipases, monoacylglycerol lipase and fatty amide hydrolase in the brain. Reductions in lysophosphatidylethanolamines (16:0; 18:0, 18:1, and 22:6) derived from the phosphatidylethanolamines and free fatty acids ( acid16:0; Linolenic acid18:3; Eicosadienoic 20:2; Arachidonic 20:4; and Docosahexaenoic 22:6) were also noted, suggesting that chlorpyrifos inhibited the of selected phospholipid signaling precursors at sublethal concentrations. These results indicate that in addition to AChE inhibition, environmentally relevant chlorpyrifos exposure alters serine lipase activity and metabolites in the trout brain, which may compromise neuronal signaling and impact neurobehavioral responses in aquatic animals.© The Author(s) 2019. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Keyword: fat metabolism

Hexadecenoic Fatty Positional Isomers and De Novo PUFA Synthesis in Colon Cancer Cells.

involves delta-9 and delta-6 desaturase enzymes forming palmitoleic (9-16:1; -7 series) and sapienic (6-16:1; -10 series), respectively. The corresponding biological consequences and lipidomic research on these positional monounsaturated fatty (MUFA) isomers are under development. Furthermore, sapienic can bring to the de novo synthesis of the -10 polyunsaturated fatty (PUFA) sebaleic (5,8-18:2), but such transformations in cancer cells are not known. The model of Caco-2 cell line was used to monitor sapienic supplementation (150 and 300 μM) and provide evidence of the formation of -10 fatty acids as well as their incorporation at levels of membrane phospholipids and triglycerides. Comparison with palmitoleic and acids evidenced that remodelling was influenced by the type of fatty and positional isomer, with an increase of 8-18:1, -10 PUFA and a decrease of saturated fats in case of sapienic . Cholesteryl esters were formed only in cases with sapienic . Sapienic was the less toxic among the tested fatty acids, showing the highest ECs and inducing death only in 75% of cells at the highest concentration tested. Two-photon fluorescent microscopy with Laurdan as a fluorescent dye provided information on membrane fluidity, highlighting that sapienic increases the distribution of fluid regions, probably connected with the formation of 8-18:1 and the -10 PUFA in cell lipidome. Our results bring evidence for MUFA positional isomers and de novo PUFA synthesis for developing lipidomic analysis and cancer research.

Keyword: fat metabolism

Adiponectin homolog novel osmotin protects obesity/diabetes-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic fatty liver disease (NAFLD) in obesity-associated diabetes.To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human liver carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum parameters, and increased fatty oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/diabetes-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

CHP1 Regulates Compartmentalized Glycerolipid Synthesis by Activating GPAT4.

Cells require a constant supply of fatty acids to survive and proliferate. Fatty acids incorporate into membrane and storage glycerolipids through a series of endoplasmic reticulum (ER) enzymes, but how these enzymes are regulated is not well understood. Here, using a combination of CRISPR-based genetic screens and unbiased lipidomics, we identified calcineurin B homologous protein 1 (CHP1) as a major regulator of ER glycerolipid synthesis. Loss of\xa0CHP1 severely reduces fatty incorporation and storage in mammalian cells and invertebrates. Mechanistically, CHP1 binds and activates GPAT4, which catalyzes the initial rate-limiting step in glycerolipid synthesis. GPAT4 activity requires CHP1 to be N-myristoylated, forming a key molecular interface between the two proteins. Interestingly, upon CHP1 loss, the peroxisomal enzyme, GNPAT, partially compensates for the loss of ER synthesis, enabling cell proliferation. Thus, our work identifies a conserved regulator of glycerolipid and reveals plasticity in synthesis of proliferating cells.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fat metabolism

Berberine Protects Glomerular Podocytes via Inhibiting Drp1-Mediated Mitochondrial Fission and Dysfunction.

Elevated levels of plasma free fatty (FFA) and disturbed mitochondrial dynamics play crucial roles in the pathogenesis of diabetic kidney disease (DKD). However, the mechanisms by which FFA leads to mitochondrial damage in glomerular podocytes of DKD and the effects of Berberine (BBR) on podocytes are not fully understood. : Using the db/db diabetic mice model and cultured mouse podocytes, we investigated the molecular mechanism of FFA-induced disturbance of mitochondrial dynamics in podocytes and testified the effects of BBR on regulating mitochondrial dysfunction, podocyte apoptosis and glomerulopathy in the progression of DKD. : Intragastric administration of BBR for 8 weeks in db/db mice significantly reversed glucose and disorders, podocyte damage, basement membrane thickening, mesangial expansion and glomerulosclerosis. BBR strongly inhibited podocyte apoptosis, increased reactive oxygen species (ROS) generation, mitochondrial fragmentation and dysfunction both and . Mechanistically, BBR could stabilize mitochondrial morphology in podocytes via abolishing (PA)-induced activation of dynamin-related protein 1 (Drp1). : Our study demonstrated for the first time that BBR may have a previously unrecognized role in protecting glomerulus and podocytes via positively regulating Drp1-mediated mitochondrial dynamics. It might serve as a novel therapeutic drug for the treatment of DKD.

Keyword: fat metabolism

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic fatty liver disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic fatty liver disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for gluconeogenesis (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in liver. Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for gluconeogenesis in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: fat metabolism

-Rich High Diet Exacerbates Experimental Pulmonary Fibrosis by Modulating Endoplasmic Reticulum Stress.

The impact of lipotoxicity on the development of lung fibrosis is unclear. Saturated fatty acids such as (PA) activate endoplasmic reticulum (ER) stress, a cellular stress response associated with the development of idiopathic pulmonary fibrosis (IPF). We tested the hypothesis that PA increases susceptibility to lung epithelial cell death and experimental fibrosis by modulating ER stress. Total liquid chromatography and mass spectrometry were used to measure fatty content in IPF lungs. Wild-type mice were fed a high diet (HFD) rich in PA or a standard diet (SD) and subjected to bleomycin-induced lung injury. Lung fibrosis was determined by hydroxyproline content. Mouse lung epithelial cells were treated with PA. ER stress and cell death were assessed by Western blotting, TUNEL staining and cell viability assays. IPF lungs had a higher level of PA compared to controls. Bleomycin-exposed mice fed a HFD had significantly increased pulmonary fibrosis associated with increased cell death and ER stress compared to those fed a SD. PA increased apoptosis and activation of the unfolded protein response in lung epithelial cells. This was attenuated by genetic deletion and chemical inhibition of CD36, a fatty transporter. In conclusion, consumption of a HFD rich in saturated increases susceptibility to lung fibrosis and ER stress and PA mediates lung epithelial cell death and ER stress via CD36. These findings demonstrate that lipotoxicity may have a significant impact on the development of lung injury and fibrosis by enhancing pro-death ER stress pathways.

Keyword: fat metabolism

-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated fatty in fast foods and is known to induce inflammation and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased body weight, hyperlipidemia, hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the liver. Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical metabolic changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic fatty liver disease.

Keyword: fat metabolism

Fatty levels alterations in THP-1 macrophages cultured with lead (Pb).

As cardiovascular events are one of the main causes of death in developed countries, each factor potentially increasing the risk of cardiovascular disease deserves special attention. One such factor is the potentially atherogenic effect of lead (Pb) on , and is significant in view of the still considerable Pb environmental pollution and the non-degradability of Pb compounds.Analysis of saturated fatty acids (SFA) (caprylic (C8:0), decanoic (C10:0), lauric (C12:0), tridecanoic (C13:0), myristic (C14:0), pentadecanoic (C15:0), (C16:0), heptadecanoic (C17:0), stearic (C18:0), and behenic (C22:0)), monounsaturated fatty (MUFA) (palmitoleic (C16:1), oleic (18:1w9), trans-vaccenic (C18:1 trans11)), and polyunsaturated fatty (PUFA) (linoleic (C18:2n6), gamma-linolenic (C18:3n6), arachidonic (C20:4n6)), was conducted by gas chromatography. Analysis of stearoyl-CoA desaturase (SCD), fatty desaturase 1 (FADS1) and fatty desaturase 2 (FADS2) expression was performed using qRT-PCR. Oxidative stress intensity (malondialdehyde - MDA concentration) was measured using spectrophotometric method. Intracellular generation of reactive oxygen species (ROS) in macrophages was visualized by fluorescence microscopy and quantitatively measured by plate reader.Pb caused quantitative alterations in FAs profile in macrophages; the effect was Pb-concentration dependent and selective (i.e. concerned only selected FAs). In general, the effect of Pb was biphasic, with Pb levels of 1.25\u2009μg/dL and 2.5\u2009μg/dL being stimulatory, and 10\u2009μg/dL being inhibitory on concentrations of selected FAs. The most potent Pb concentration, resulting in increase in levels of 9 FAs, was 2.5\u2009μg/dL, the Pb-level corresponding to the mean blood Pb concentrations of people living in urban areas not contaminated by Pb. Pb was found to exert similar, biphasic effect on the expression of FADS1. However, Pb decreased, in a concentration-dependent manner, the expression of SCD and FADS2. Pb significantly increased MDA and ROS concentration in macrophages.Environmental Pb exposure might be a risk factor resulting in alterations in FAs levels, oxidative stress and increased MDA concentration in macrophages, which might lead to the formation of foam cells and to inflammatory reactions.Copyright © 2019 Elsevier GmbH. All rights reserved.

Keyword: fat metabolism

Carbon monoxide releasing molecule-A1 improves nonalcoholic steatohepatitis via Nrf2 activation mediated improvement in oxidative stress and mitochondrial function.

Nuclear factor-erythroid 2 related factor 2 (Nrf2)-mediated signaling plays a central role in maintaining cellular redox homeostasis of hepatic cells. Carbon monoxide releasing molecule-A1 (CORM-A1) has been reported to stimulate up-regulation and nuclear translocation of Nrf2 in hepatocytes. However, the role of CORM-A1 in improving , antioxidant signaling and mitochondrial functions in nonalcoholic steatohepatitis (NASH) is unknown. In this study, we report that CORM-A1 prevents hepatic steatosis in high high fructose (HFHF) diet fed C57BL/6J mice, used as model of NASH. The beneficial effects of CORM-A1 in HFHF fed mice was associated with improved homeostasis, Nrf2 activation, upregulation of antioxidant responsive (ARE) genes and increased ATP production. As, mitochondria are intracellular source of reactive oxygen species (ROS) and important sites of , we further investigated the mechanisms of action of CORM-A1-mediated improvement in mitochondrial function in (PA) treated HepG2 cells. Cellular oxidative stress and cell viability were found to be improved in PA\xa0+\xa0CORM-A1 treated cells via Nrf2 translocation and activation of cytoprotective genes. Furthermore, in PA treated cells, CORM-A1 improved mitochondrial oxidative stress, membrane potential and rescued mitochondrial biogenesis thru upregulation of Drp1, TFAM, PGC-1α and NRF-1 genes. CORM-A1 treatment improved cellular status by lowering glycolytic respiration and maximizing OCR. Improvement in mitochondrial respiration and increment in ATP production in PA\xa0+\xa0CORM-A1 treated cells further corroborate our findings. In summary, our data demonstrate for the first time that CORM-A1 ameliorates tissue damage in steatotic liver via Nrf2 activation and improved mitochondrial function, thus, suggesting the anti-NASH potential of CORM-A1.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: fat metabolism

Mutation of IDH1 aggravates the fatty ‑induced oxidative stress in HCT116 cells by affecting the mitochondrial respiratory chain.

Increasing evidence has indicated that mutations of isocitrate dehydrogenase 1/2 (IDH1/2) contribute to the metabolic reprogramming of cancer cells; however their functions in remain unknown. In the present study, the parental and IDH1 (R132H/+) mutant HCT116 cells were treated with various concentrations of oleic (OA) or (PA) in the presence or absence of glucose. The results demonstrated that mutation of IDH1 exacerbated the effects of OA and PA on cell viability and apoptosis, and consistently elevated the production of reactive oxygen species in HCT116 cells, particularly in the absence of glucose. Furthermore, mutation of IDH1 inhibited the rate of fatty oxidation (FAO), but elevated the glucose consumption in HCT116 cells. The results of immunoblotting and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) indicated that the expression of glucose transporter 1 was upregulated, whereas that of carnitine palmitoyl transferase 1 was downregulated in IDH1 mutant HCT116 cells. Although mitochondrial DNA quantification demonstrated that mutation of IDH1 had no effect on the quantity of mitochondria, immunoblotting and RT‑qPCR revealed that mutation of IDH1 in HCT116 cells significantly downregulated the expression of cytochrome c (CYCS) and CYCS oxidase IV, two important components in mitochondrial respiratory chain. These results indicated that mutation of IDH1 aggravated the fatty ‑induced oxidative stress in HCT116 cells, by suppressing FAO and disrupting the mitochondrial respiratory chain. The results of the present study may provide novel insight into therapeutic strategies for the treatment of cancer types with IDH mutation.

Keyword: fat metabolism

Apigenin reduces the excessive accumulation of lipids induced by via the AMPK signaling pathway in HepG2 cells.

In recent years, increasing attention has been paid to diseases caused by excessive accumulation of lipids in the liver with therapeutic agents derived from natural products offering an alternative treatment to conventional therapies. Among these therapeutic agents, apigenin, a natural flavonoid, has been proven to exert various beneficial biological effects. In the present study, the antiadipogenic effects of apigenin in HepG2 cells was investigated. It was demonstrated that the treatment of cells with different concentrations of apigenin for 24 h significantly decreased the -induced increases in total cholesterol (TC) and triglyceride (TG) levels as well as intracellular accumulation. In addition, apigenin increased the phosphorylated-AMP-activated protein kinase (AMPK) levels but decreased the expression levels of 3-hydroxy-3-methylglutaryl CoA reductase, sterol regulatory element-binding protein (SREBP)-1, fatty synthase, and SREBP-2 in a concentration-dependent manner. The present findings suggested that apigenin might improve by activating the AMPK/SREBP pathway to reduce accumulation in the liver.Copyright: © Lu et al.

Keyword: fat metabolism

[Comparison of effects of oleic and on deposition and mTOR / S6K1 / SREBP-1c pathway in HepG2 cells].

To explore the effects of oleic and on deposition and mTOR/S6K1/SREBP-1c pathways in HepG2 cells. The model of steatosis was established with induction of oleic and and was intervened by rapamycin. The changes in droplets were observed after staining the cells with oil Red O. Intracellular triglyceride (TG) contents in cells were measured by TG kit. mTOR, S6K1, and SREBP-1c mRNA expression levels were detected using QRT-PCR. Western blot was used to determine protein expression levels of mTOR, S6K1 and SREBP-1c. Both fatty acids increased droplets in HepG2 cells. Fatty degeneration with elevated TG occurred with significant changes in oleic group lipids. Rapamycin alleviated deposition caused by oleic and and inhibited their induction of increased expression of mTOR, S6K1, and SREBP-1c. QRT-PCR and Western blot results showed that mRNA and protein expressions of mTOR, S6K1, and SREBP-1c in oleic and group were significantly higher than the control group ( < 0.05). The increase was more pronounced in the group ( < 0.05); however, after rapamycin intervention, the expression of mRNA and protein in the three groups were significantly lower ( < 0.05), and the change in group was more pronounced ( < 0.05). Oleic and can induce deposition in HepG2 cells and increase expression of every component of mTOR/S6K1/SREBP-1c pathway; however, Oleic -induced deposition is more pronounced, and the mTOR, S6K1, and SREBP-1c pathway change is more obvious in . Rapamycin has high potent inhibitory effect on -induced deposition. These results specify that synthesis involved in the mTOR/S6K1/SREBP-1c pathways are mainly associated to in HepG2 cells, whereas other signaling pathway may mediate oleic -induced synthesis.

Keyword: fat metabolism

The effect of saturated and unsaturated acids on the production of outer membrane vesicles from and .

The aim of present study is to investigate the effect of acids on the outer membrane vesicles (OMVs) produced by spp. spp. is the important member of Gut microbiota that employ OMVs production for interact with host. Besides, dietary acids could influence on determination of gut microbiota composition and immune response. In this regard, we evaluated the effect of acids on the growth and OMVs production of and . and were grown on BHI broth with and without and palmitoleic acids as saturated and unsaturated acids, respectively. OMVs were extracted using multiple centrifugation and tris-ethylene diamine tetra acetic (EDTA)-Sodium deoxy cholate buffers. Physicochemical properties of OMVs were detected by electron microscopy (SEM), Bradford Coomassie brilliant blue assay and SDS-PAGE. Data were analyzed with One-way ANOVA using SPSS.The growths of both were significantly increased by . Nevertheless, palmitoleic had no significant effect on them. significantly decreased and increased the production of OMVs at low and high concentration, respectively. However, the production of OMVs was not significantly affected by . Although palmitoleic had a significant decreasing effect on the production of OMVs, it significantly increased the production of OMVs at low concentration.In conclusion we reported that had a stimulatory effect on the growth of and and had a dose dependent effect on the production of OMVs. Also producing of OMVs was affected by palmitoleic in a dose dependent manner.

Keyword: fatty liver

Phytochemistry, Traditional Uses and Pharmacological Profile of Rose Hip: A Review.

The fruit of genus Rosa, known as "rose hip", is frequently used in different traditional medicines. Rose hips have long been used to treat kidney stones, gastroenteric ailments, hypertension and respiratory problems such as bronchitis, cough and cold.This review is focused on the ethnopharmacological uses of rose hip as well as phytochemical and pharmacological aspects.Ethno-medical uses of rose hip have been recorded in many countries since a long time. Approximately, 129 chemical compounds have been isolated and identified from rose hip. This fruit contains some major active components such as flavonoids, tannins, anthocyanin, phenolic compounds, oil, organic acids and inorganic compounds. Scientific studies have suggested a wide range of pharmacological activities for rose hip including antioxidant, anti-inflammatory, anti-obesity, anti-cancer, hepatoprotective, nephroprotective, cardioprotective, antiaging, anti H. pylori, neuroprotective and antinociceptive activities. In particular, the rose hip powder and extract have been reported to exert therapeutic effects on arthritis.Some of the ethnomedical indications of rose hip, such as nephroprotective and gastroproetctive actions, have been confirmed by preclinical pharmacological studies. Additional investigations on the pharmacological effects of rose hip as well as evidence from randomized controlled trials are essential to assess the therapeutic value of this natural product.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keyword: fatty liver

Trigonelline prevents high cholesterol and high fat diet induced hepatic lipid accumulation and lipo-toxicity in C57BL/6J mice, via restoration of hepatic autophagy.

Non-alcoholic disease (NAFLD) is often linked with impaired hepatic autophagy. Here, we studied the alterations in hepatocellular autophagy by high cholesterol and high-fat diet (HC-HF) diet in C57BL/6J mice, and by (PA), in AML-12 and HepG2 cells. Further, we analysed role of Trigonelline (TG), a plant alkaloid, in preventing NAFLD, by modulating autophagy. For this, C57BL/6J mice were fed with Standard Chow (SC) or HC-HF diet, with and without TG for 16 weeks. In-vitro; AML-12\u202fcells and HepG2 cells, were exposed to PA with and without TG, for 24\u202fh. Cellular events related to autophagy, lipogenesis, and lipo-toxicity were studied. The HC-HF diet fed mice showed hepatic autophagy blockade, increased triglycerides and steatosis. PA exposure to AML-12\u202fcells and HepG2 cells induced impaired autophagy, ER stress, resulting in lipotoxicity. TG treatment in HC-HF fed mice, restored hepatic autophagy, and prevented steatosis. TG treated AML-12, and HepG2 cells exposed to PA showed autophagy restoration, and reduced lipotoxicity, however, these effects were diminished in Atg7-/- HepG2 cells, and in the presence of chloroquine. This study shows that HC-HF diet-induced impaired autophagy, and steatosis is prevented by TG, which attributes to its novel mechanism in treating NAFLD.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Nobiletin alleviates ‑induced NLRP3 inflammasome activation in a sirtuin 1‑dependent manner in AML‑12 cells.

The NOD‑like receptor family pyrin domain containing 3 (NLRP3) inflammasome has been reported to contribute to (PA)‑induced lipotoxicity. Nobiletin (Nob) is a polymethoxylated flavonoid derived from citrus fruits that has been reported to exert antioxidant and antitumor effects. However, its protective and regulatory mechanisms in PA‑induced lipotoxicity remain unclear. Therefore, the aim of the present study was to investigate the protective effects of Nob in AML‑12 cells against lipotoxicity and examine the underlying mechanism. Western blotting, reverse transcription‑quantitative polymerase chain reaction and ELISA assays were performed to investigate the activation of the NLRP3 inflammasome. Sirtuin 1 (SIRT1) small interfering RNA was used to knockdown SIRT1 expression in AML‑12 cells. The results demonstrated that PA effectively activated NLRP3 inflammasome and increased the expression and secretion of interleukin (IL)‑1β and IL‑18. Notably, the PA‑induced inflammasome activation was reversed by Nob, as indicated by the decreased expression levels of NLRP3, Caspase‑1, IL‑1β and IL‑18. Furthermore, Nob treatment with or without PA enhanced the expression of SIRT1 in AML‑12 cells, while knockdown of SIRT1 with SIRT1‑small interfering RNA reversed the anti‑inflammatory effects of Nob. Overall, the results of the present study indicated that Nob alleviated PA‑induced lipotoxicity in AML‑12 cells via the suppression of NLRP3 inflammasome activation in a SIRT1‑dependent manner. These results provide a possible basis of the underlying mechanism and, in turn, the potential application of Nob in the treatment of non‑alcoholic disease.

Keyword: fatty liver

Development of an in vitro model to study hepatitis C virus effects on hepatocellular lipotoxicity and lipid metabolism.

Hepatic steatosis is common in patients infected with hepatitis C virus (HCV). Particularly in patients infected with non-genotype 3 HCV, hepatic steatosis is closely related to factors of the metabolic syndrome such as hyperlipidemia. However, the molecular mechanisms involved in this "metabolic" steatosis in non-3 genotype HCV infections are not well understood. Here, we aimed to develop an in vitro model to study the effect of genotype 1 HCV infection on hepatic lipotoxicity and lipid metabolism. Cellular lipid accumulation was induced in Huh-7 hepatoma cells transfected with HCV genotype 1b replicon (HCV) by incubation with increasing doses of (C16:0) or oleic (C18:1 n-9) complexed to albumin mimicking hyperlipidemic conditions. Mock transfected hepatoma cells (HCV) were used as controls. Incubation with oleic concentrations as high as 0.5\u202fmM did not induce toxic effects in HCV or HCV cells. In contrast, incubation with caused dose-dependently cytotoxic effects which were more pronounced in HCV compared to HCV cells. Further analysis with subtoxic and oleic concentrations revealed a higher uptake of acids and intracellular triglyceride accumulation in HCV compared to HCV cells. Carnitine palmitoyltransferase I (CPT1) expression, indicative of mitochondrial beta-oxidation, was markedly stimulated by lipid exposure in HCV but not in HCV cells. Furthermore, heme oxygenase 1 (HMOX1) expression levels increased in FA stimulated cells, and this increase was significantly higher in HCV compared to HCV cells. In contrast, expression of the key enzymes of hepatic de novo lipogenesis synthase (FASN) and stearoyl-CoA desaturase (SCD-1) was significantly reduced upon oleate exposure in HCV but not in HCV cells. In summary, our newly developed cell culture model revealed effects of HCV genotype 1b infection on metabolic susceptibility to lipid accumulation and toxicity particularly to saturated lipids. These results may indicate that HCV (genotype 1b) infected individuals with hyperlipidemia may benefit from dietary or pharmacological intervention.Copyright © 2018 Elsevier GmbH. All rights reserved.

Keyword: fatty liver

Oleate ameliorates palmitate-induced reduction of NAMPT activity and NAD levels in primary human hepatocytes and hepatocarcinoma cells.

Nicotinamide phosphoribosyltransferase (NAMPT) and nicotinamide adenine dinucleotide (NAD) levels are crucial for function. The saturated palmitate and the unsaturated oleate are the main free acids in adipose tissue and human diet. We asked how these acids affect cell survival, NAMPT and NAD levels in HepG2 cells and primary human hepatocytes.HepG2 cells were stimulated with palmitate (0.5mM), oleate (1mM) or a combination of both (0.5mM/1mM) as well as nicotinamide mononucleotide (NMN) (0.5 mM) or the specific NAMPT inhibitor FK866 (10nM). Cell survival was measured by WST-1 assay and Annexin V/propidium iodide staining. NAD levels were determined by NAD/NADH Assay or HPLC. Protein and mRNA levels were analysed by Western blot analyses and qPCR, respectively. NAMPT enzyme activity was measured using radiolabelled C-nicotinamide. Lipids were stained by Oil red O staining.Palmitate significantly reduced cell survival and induced apoptosis at physiological doses. NAMPT activity and NAD levels significantly declined after 48h of palmitate. In addition, NAMPT mRNA expression was enhanced which was associated with increased NAMPT release into the supernatant, while intracellular NAMPT protein levels remained stable. Oleate alone did not influence cell viability and NAMPT activity but ameliorated the negative impact of palmitate on cell survival, NAMPT activity and NAD levels, as well as the increased NAMPT mRNA expression and secretion. NMN was able to normalize intracellular NAD levels but did not ameliorate cell viability after co-stimulation with palmitate. FK866, a specific NAMPT inhibitor did not influence lipid accumulation after oleate-treatment.Palmitate targets NAMPT activity with a consequent cellular depletion of NAD. Oleate protects from palmitate-induced apoptosis and variation of NAMPT and NAD levels. Palmitate-induced cell stress leads to an increase of NAMPT mRNA and accumulation in the supernatant. However, the proapoptotic action of palmitate seems not to be mediated by decreased NAD levels.

Keyword: fatty liver

Differential capability of metabolic substrates to promote hepatocellular lipid accumulation.

Excessive storage of triacylglycerides (TAGs) in lipid droplets within hepatocytes is a hallmark of non-alcoholic disease (NAFLD), one of the most widespread metabolic disorders in Western societies. For the purpose of exploring molecular pathways in NAFLD development and testing potential drug candidates, well-characterised experimental models of ectopic TAG storage in hepatocytes are needed.Using an optimised Oil Red O assay, immunoblotting and real-time qRT-PCR, we compared the capability of dietary monosaccharides and acids to promote lipid accumulation in HepG2 human hepatoma cells.Both high glucose and high fructose resulted in intracellular lipid accumulation after 48\xa0h, and this was further augmented (up to twofold, as compared to basal levels) by co-treatment with the lipogenesis-stimulating hormone insulin and the pro-inflammatory cytokine tumour necrosis factor alpha (TNF-α), respectively. The acids and oleic were even more effective than these carbohydrates, inducing significantly elevated TAG storage already after 24\xa0h of treatment. Highest (about threefold) increases in lipid accumulation were observed upon treatment with oleic , alone as well as in combinations with or with high glucose and insulin. Increases in protein levels of a major lipid droplet coat protein, perilipin-2 (PLIN2), mirrored intracellular lipid accumulation following different treatment regimens.Several treatment regimens of excessive fat and sugar supply promoted lipid accumulation in HepG2 cells, albeit with differences in the extent and rapidity of steatogenesis. PLIN2 is a candidate molecular marker of sustained lipid accumulation in HepG2 cells.

Keyword: fatty liver

Fermented Soymilk Alleviates Lipid Accumulation by Inhibition of SREBP-1 and Activation of NRF-2 in the Hepatocellular Steatosis Model.

Ingredients of soy and fermented soy products have been widely utilized as food supplements for health-enhancing properties. The aim of this study was to evaluate the effects of fermented soymilk (FSM) and soymilk (SM) on free -induced lipogenesis in the hepatocellular steatosis model. HepG2 cells were incubated with (PA) for 24 h to induce lipogenesis and accumulation of intracellular lipid contents. The PA-treated cells were co-incubated with FSM, SM, genistein, and estrogen, respectively. Lipid accumulation in the PA-treated HpG2 cells was significantly decreased by co-incubation with FSM. Treatment of HepG2 cells with PA combined with genistein or estrogen significantly increased the expression of SREBP-1. However, FSM co-incubation significantly attenuated SREBP-1 expression in the PA-treated HepG2 cells; in addition, expression of NRF-2 and phosphorylation of ERK were significantly increased in the PA and FSM co-incubated cells. PA-induced ROS production was significantly reduced by FSM and SM. Our results suggested that the bioactive components of FSM could protect hepatocytes against the lipid accumulation and ROS production induced by free acids. These effects may be mediated by the inhibition of SREBP-1 and the activation of NRF-2 via the ERK pathway in HepG2 cells.

Keyword: fatty liver

Compound C Protects Mice from HFD-Induced Obesity and Nonalcoholic Disease.

The aim of this study was to investigate the effects of compound C on an in vivo mouse model of high-fat diet- (HFD-) induced obesity and hepatosteatosis.C57BL/6 mice were fed with a standard diet (\u2009=\u20095) for 16\u2009weeks and then injected saline once a day for 4\u2009weeks as the normal chow group. Mice (\u2009=\u200910) were fed with HFD for 16\u2009weeks to induce obesity and hepatosteatosis and then divided into two groups: HFD\u2009+\u2009vehicle group injected with the vehicle solution (saline) and HFD\u2009+\u2009compound C group injected with compound C in saline (5\u2009mg/kg i.p., once a day) for 4\u2009weeks. histology was observed. The expression levels of genes related to lipid metabolism and proinflammation in tissue were examined. NLRP3 inflammasome expression in tissue was detected by the western blot assay. HepG2 cells were pretreated with compound C and/or AICAR for 1\u2009h and then treated with (PA) for 3\u2009h. The cells were collected, and mRNA levels were determined.There was a significant reduction in body-weight gain and daily food intake in the HFD\u2009+\u2009compound C group compared with the HFD\u2009+\u2009vehicle group ( < 0.05). The glucose tolerance test (GTT) and insulin tolerance test (ITT) showed that compound C alleviated insulin resistance. Histology analysis showed a significant reduction of hepatic steatosis by compound C. Compound C also significantly decreased synthesis genes, while increased oxidation genes. Furthermore, compound C significantly reduced the expression of proinflammatory markers and NLRP3 inflammasome ( < 0.05). Compound C enhanced mRNA levels of SOD1, SOD2, catalase, GPx1, and GPx4 and reduced the p-AMPK/AMPK ratio, which were stimulated by (PA). The effect was enhanced by AICAR.Our data suggest that compound C is a potent NAFLD suppressor and an attractive therapeutic target for hepatic steatosis and related metabolic disorders.

Keyword: fatty liver

Does the enterolactone (ENL) affect transporters and lipid metabolism in ?

NAFLD as a result of inappropriate diet and obesity, may progress to sever conditions such as: type 2 diabetes mellitus or steatohepatitis, and has recently become a prevalent topic of numerous investigations. Due to its dangerous aftermaths, finding new substances, such as polyphenols and their derivatives, which might reduce steatosis is the main target of research into NAFLD treatment. Hence, the aim of the present study was to evaluate the effect(s) of enterolactone (ENL), a metabolite of secoisolariciresinol (SECO), on lipid metabolism together with changes in the expression of transporters in .The experiments were conducted on HepG2 cells incubated with either ENL and/or during 16\xa0h exposure. The expression of selected transport proteins: FATP2, FATP5, CD36, FABPpm, ABCA1, MTP, ACBP and L-FABP, as well as the proteins directly involved in lipogenesis (FAS), oxidation pathway (CPT 1), and lipid metabolism (PPARα, LXR, SREBP1c, pAMPK) was estimated by Western Blot. Intra and extracellular lipid contents were assessed by Gas-Liquid Chromatography. The data was analyzed with two-way analysis of variance (ANOVA), and results were considered to be statistically significant at \xa0≤\xa00.05.ENL stimulated extracellular efflux of free acids (FFA) and triacylglicerols (TAG) to the medium, while, it had no influence on FATP-family mediated intracellular uptake. Moreover, ENL decreased the expression of CPT 1, pAMPK, PPARα, increased SREBP1c and had no effect on LXR, and FAS content.The findings of our study demonstrate that ENL had opposite effect on steatosis in comparison with other polyphenols what suggests that it may be an inactive metabolite. ENL did not affect significantly the intracellular accumulation of FFA, DAG and TAG, yet it promoted their extracellular efflux. Furthermore, it inhibited ß-oxydation and intracellular lipid metabolism what may contribute to the progression of NAFLD.

Keyword: fatty liver

Membranotropic effects of ω-hydroxypalmitic and Ca on rat mitochondria and lecithin liposomes. Aggregation and membrane permeabilization.

The paper examines membranotropic Ca-dependent effects of ω-hydroxypalmitic (HPA), a product of ω-oxidation of acids, on the isolated rat mitochondria and artificial membrane systems (liposomes). It was established that in the presence of Ca, HPA induced aggregation of mitochondria, which was accompanied by the release of cytochrome c from the organelles. It was further demonstrated that the addition of Ca to HPA-containing liposomes induced their aggregation and/or fusion. Ca also caused the release of the fluorescent dye sulforhodamine B from liposomes, indicating their permeabilization. HPA was shown to induce a high-amplitude swelling of Ca-loaded mitochondria, to decrease their membrane potential, to induce the release of Ca from the organelles and to result in the oxidation of the mitochondrial NAD(P)H pool. Those effects of HPA were not blocked by the MPT pore inhibitor CsA, but were suppressed by the mitochondrial calcium uniporter inhibitor ruthenium red. The effects of HPA were also observed when Ca was replaced with Sr (but not with Ba or Mg). A supposition is made that HPA can induce a Ca-dependent aggregation of mitochondria, as well as Cadependent CsA-insensitive permeabilization of the inner mitochondrial membrane - with the subsequent lysis of the organelles.

Keyword: fatty liver

Ginsenoside Rg1 Ameliorates -Induced Hepatic Steatosis and Inflammation in HepG2 Cells via the AMPK/NF-B Pathway.

Nonalcoholic disease (NAFLD) is one of the common diseases in the world, and it can progress from simple lipid accumulation to sustained inflammation. The present study was designed to investigate the effects and underlying mechanisms of ginsenoside Rg1 (G-Rg1) treatment on NAFLD . HepG2 cells were treated with (PA) to induce steatosis and inflammation and then successively incubated with G-Rg1. Lipids accumulation was analyzed by Oil Red O staining and intracellular triglyceride (TG) quantification. Inflammatory conditions were examined by quantifying the levels of cell supernatant alanine transaminase/aspartate aminotransferase (ALT/AST) and secretory proinflammatory cytokines, including IL-1, IL-6, and TNF- in the cell supernatants. Quantitative RT-PCR and western blotting were used to measure the expressions of genes and proteins associated with lipogenic synthesis and inflammation, including AMP-activated protein kinase (AMPK) and nuclear factor-kappa B (NF-B) pathways. HepG2 cells were pretreated with an AMPK inhibitor; then, Oil Red O staining and TG quantification were performed to study the lipid deposition. Phospho-AMPK (Thr172) (p-AMPK) and phospho-acetyl-CoA carboxylase (Ser79) (p-ACC) were quantified by immunoblotting. Immunofluorescence was performed to demonstrate the nuclear translocation of NF-B P65. The present study showed that PA markedly increased the intracellular lipid droplets accumulation and TG levels, but decreased AMPK phosphorylation and the expressions of its downstream lipogenic genes. However, G-Rg1 alleviated hepatic steatosis and reduced the intracellular TG content; these changes were accompanied by the activation of the AMPK pathway. In addition, blocking AMPK by using the AMPK inhibitor markedly abolished the G-Rg1-mediated protection against PA-induced lipid deposition in HepG2 cells. Furthermore, G-Rg1 reduced the ALT/AST levels and proinflammatory cytokines release, which were all enhanced by PA. These effects were correlated with the inactivation of the NF-B pathway and translocation of P65 from the cytoplasm to the nucleus. Overall, these results suggest that G-Rg1 effectively ameliorates hepatic steatosis and inflammation, which might be associated with the AMPK/NF-B pathway.

Keyword: fatty liver

Dual roles of ULK1 (unc-51 like autophagy activating kinase 1) in cytoprotection against lipotoxicity.

Saturated (SFA)-induced lipotoxicity is caused by the accumulation of reactive oxygen species (ROS), which is associated with damaged mitochondria. Moreover, lipotoxicity is crucial for the progression of nonalcoholic steatohepatitis (NASH). Autophagy is required for the clearance of protein aggregates or damaged mitochondria to maintain cellular metabolic homeostasis. The NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2)-KEAP1 (kelch like ECH associated protein 1) pathway is essential for the elimination of ROS. ULK1 (unc-51 like autophagy activating kinase 1; yeast Atg1) is involved in the initiation of autophagy; however, its role in lipotoxicity-induced cell death in hepatocytes and mouse has not been elucidated. We now show that ULK1 potentiates the interaction between KEAP1 and the autophagy adaptor protein SQSTM1/p62, thereby mediating NFE2L2 activation in a manner requiring SQSTM1-dependent autophagic KEAP1 degradation. Furthermore, ULK1 is required for the autophagic removal of damaged mitochondria and to enhance binding between SQSTM1 and PINK1 (PTEN induced kinase 1). This study demonstrates the molecular mechanisms underlying the cytoprotective role of ULK1 against lipotoxicity. Thus, ULK1 could represent a potential therapeutic target for the treatment of NASH. Abbreviations: ACTB: actin beta; CM-HDCFDA:5-(and-6)-chloromethyl-2\',7\'-dichlorodihydrofluorescein diacetate; CQ: chloroquine; CUL3: cullin 3; DMSO: dimethyl sulfoxide; GSTA1: glutathione S-transferase A1; HA: hemagglutinin; Hepa1c1c7: mouse hepatoma cells; HMOX1/HO-1: heme oxygenase 1; KEAP1: kelch like ECH associated protein 1; LPS: lipopolysaccharides; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MAPK8/JNK: mitogen-activated protein kinase 8; MEF: mouse embryonic fibroblast; MFN1: mitofusin 1; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NASH: nonalcoholic steatohepatitis; NFE2L2/NRF2: nuclear factor, erythroid 2 like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PA: ; PARP: poly (ADP-ribose) polymerase 1; PINK1: PTEN induced kinase 1; PRKAA1/2: protein kinase AMP-activated catalytic subunits alpha1/2; PRKN/PARK2: parkin RBR E3 ubiquitin protein ligase; PRKC/PKC: protein kinase C; RBX1: ring-box 1; ROS: reactive oxygen species; SFA: saturated ; siRNA: small interfering RNA; SQSTM1/p62: sequestosome 1; TOMM20: translocase of outer mitochondrial membrane 20; TUBA: tubulin alpha; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; ULK1: unc-51 like autophagy activating kinase 1.

Keyword: fatty liver

Beyond the Scavenging of Reactive Oxygen Species (ROS): Direct Effect of Cerium Oxide Nanoparticles in Reducing Acids Content in an In Vitro Model of Hepatocellular Steatosis.

Nonalcoholic disease (NAFLD) is characterized by hepatic accumulation of lipids. Antisteatotic effects of cerium oxide nanoparticles (CeONPs) have recently been shown in animal models of disease. However, it is unclear whether the activity of CeONPs is related solely to the decrease in oxidative stress or, in addition, they directly decrease accumulation. To address this question, in this work, we used an in vitro model of hepatocellular steatosis, exposing HepG2 cells to oleic and . Cell uptake of CeONPs and their effect on oxidative stress and viability of hepatic cells cultured with HO were also evaluated. Results show that CeONPs were uptaken by HepG2 cells and reduced oxidative stress and improved cell viability. Treatment with oleic and increased lipogenesis and the content of different acids. CeONPs reduced and stearic and most acids consisting of more than 18 carbon atoms. These effects were associated with significant changes in elongase and desaturase activity. In conclusion, CeONPs directly protected HepG2 cells from cell injury in oxidative stress conditions and reduced content in steatotic conditions by inducing specific changes in metabolism, thus showing potential in the treatment of NAFLD.

Keyword: fatty liver

Flavone glycosides from Sicyos angulatus and their inhibitory effects on hepatic lipid accumulation.

A library of extracted natural materials (Korea Bioactive Natural Material Bank) have been screened to discover candidates for the treatment of non-alcoholic disease (NAFLD), and the 70% ethanol extract of Sicyos angulatus was found to inhibit hepatic lipid accumulation. Bioassay-guided fractionation of this bioactive extract yielded five previously undescribed flavonoid glycosides and one previously undescribed flavonolignan glycoside along with seven known flavonoid glycosides. The chemical structures of these compounds were elucidated by a combination of extensive spectroscopic analysis, including MS, NMR and UV techniques. Eight compounds of all isolated compounds showed inhibitory effects on the lipid accumulation induced by high concentrations of and glucose in HepG2 cells. Four selected compounds were tested for lipid content in a dose-dependent manner (10, 20 and 40\u202fμM), and among those compounds, kaempferol 3-O-β-d-glucopyranosyl-7-O-α-l-rhamnopyranoside showed the strongest inhibition of hepatic lipid production in HepG2 cells. In an oil-red O staining assay, five compounds were shown to reduce hepatic lipid accumulation better than what was observed in the vehicle control group. The present study suggests a new class of chemical entities for developing bioactive agents for the treatment of diseases caused by fat accumulation in the .Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Picroside II attenuates accumulation in HepG2 cells via modulation of uptake and synthesis.

Hepatic steatosis is caused by an imbalance between free acids (FFAs) uptake, utilization, storage, and disposal. Understanding the molecular mechanisms involved in FFAs accumulation and its modulation could drive the development of potential therapies for Nonalcoholic disease. The aim of the current study was to explore the effects of picroside II, a phytoactive found in , on accumulation vis-à-vis silibinin, a known hepatoprotective phytoactive from .HepG2 cells were loaded with FFAs (oleic :/2:1) for 20 hours to mimic hepatic steatosis. The FFAs concentration achieving maximum fat accumulation and minimal cytotoxicity (500 μM) was standardized. HepG2 cells were exposed to the standardized FFAs concentration with and without picroside II pretreatment.Picroside II pretreatment inhibited FFAs-induced lipid accumulation by attenuating the expression of transport protein 5, sterol regulatory element binding protein 1 and stearoyl CoA desaturase. Preatreatment with picroside II was also found to decrease the expression of forkhead box protein O1 and phosphoenolpyruvate carboxykinase.These findings suggest that picroside II effectively attenuated accumulation by decreasing FFAs uptake and lipogenesis. Picroside II also decreased the expression of gluconeogenic genes.

Keyword: fatty liver

GCN2 deficiency protects against high fat diet induced hepatic steatosis and insulin resistance in mice.

Nonalcoholic disease (NAFLD) is characterized by hepatic lipid deposition and oxidative stress. It has been demonstrated that general control nonderepressible 2 (GCN2) is required to maintain hepatic homeostasis under conditions of amino deprivation. However, the impact of GCN2 on the development of NAFLD has not been investigated. In this study, we used Gcn2 mice to investigate the effect of GCN2 on high fat diet (HFD)-induced hepatic steatosis. After HFD feeding for 12\u202fweeks, Gcn2 mice were less obese than wild-type (WT) mice, and Gcn2 significantly attenuated HFD-induced dysfunction, hepatic steatosis and insulin resistance. In the livers of the HFD-fed mice, GCN2 deficiency resulted in higher levels of lipolysis genes, lower expression of genes related to FA synthesis, transport and lipogenesis, and less induction of oxidative stress. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, -induced steatosis, oxidative & ER stress, and changes of peroxisome proliferator-activated receptor gamma (PPARγ), synthase (FAS) and metallothionein (MT) expression in HepG2 cells. Collectively, our data provide evidences that GCN2 deficiency protects against HFD-induced hepatic steatosis by inhibiting lipogenesis and reducing oxidative stress. Our findings suggest that strategies to inhibit GCN2 activity in the may provide a novel approach to attenuate NAFLD development.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Docosahexaenoic Reduces -Induced Endoplasmic Reticulum Stress in Pancreatic Β Cells.

Endoplasmic reticulum (ER) stress leads to peripheral insulin resistance and the progression of pancreatic beta cell failure in type 2 diabetes. Although ER stress plays an important role in the pathogenesis of diabetes, it is indispensable for cellular activity. Therefore, when assessing the pathological significance of ER stress, it is important to monitor and quantify ER stress levels. Here, we have established a novel system to monitor ER stress levels quickly and sensitively, and using this method, we have clarified the effect of differences in glucose concentration and various acids on the ER of pancreatic β cells. First, we developed a cell system that secretes Gaussia luciferase in culture medium depending on the activation of the GRP78 promoter. This system could sensitively monitor ER stress levels that could not be detected with real-time RT-PCR and immunoblotting. This system revealed that hyperglycemia does not induce unfolded protein response (UPR) in a short period of time in MIN6 cells, a mouse pancreatic β cell line. Physiological concentrations of , a saturated , induced ER stress quickly, while physiological concentrations of oleic , an unsaturated , did not. Docosahexaenoic , an n-3 unsaturated , inhibited -induced ER stress. In this study, we have established a system that can sensitively detect ER stress levels of living cells in a short period of time. This system can be used to monitor the state of the ER in living cells and lead to the investigation of the significance of physiological or pathological ER stress levels.

Keyword: fatty liver

The NOX1 isoform of NADPH oxidase is involved in dysfunction of sinusoids in nonalcoholic disease.

The increased production of reactive oxygen species (ROS) has been postulated to play a key role in the progression of nonalcoholic disease (NAFLD). However, the source of ROS and mechanisms underlying the development of NAFLD have yet to be established. We observed a significant up-regulation of a minor isoform of NADPH oxidase, NOX1, in the of nonalcoholic steatohepatitis (NASH) patients as well as of mice fed a high-fat and high-cholesterol (HFC) diet for 8 weeks. In mice deficient in Nox1 (Nox1KO), increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 demonstrated in HFC diet-fed wild-type mice (WT) were significantly attenuated. Concomitantly, increased protein nitrotyrosine adducts, a marker of peroxynitrite-induced injury detected in hepatic sinusoids of WT, were significantly suppressed in Nox1KO. The expression of NOX1 mRNA was much higher in the fractions of enriched sinusoidal endothelial cells (LSECs) than in those of hepatocytes. In primary cultured LSECs, (PA) up-regulated the mRNA level of NOX1, but not of NOX2 or NOX4. The production of nitric oxide by LSECs was significantly attenuated by PA-treatment in WT but not in Nox1KO. When the in vitro relaxation of TWNT1, a cell line that originated from hepatic stellate cells, was assessed by the gel contraction assay, the relaxation of stellate cells induced by LSECs was attenuated by PA treatment. In contrast, the relaxation effect of LSECs was preserved in cells isolated from Nox1KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impaired hepatic microcirculation through the reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the progression of NAFLD.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

[Ozone oxidizes oleic with the highest rate constant and does not oxidize . Different physicochemical parameters of substrates and their role in phylogenesis.]

Physicochemical differences between О3 oxidation parameters for and oleic acids (FA) during phylogenesis (evolution) are fundamental for а) production of palmitoleic monounsaturated (MFA), b) formation of carnitine palmitoyltransferase as a FA transporter to mitochondria, and c) in vivo production of oleic MFA under humoral regulatory effect of insulin. In the strive for the best kinetic parameters of biological organisms without a possibility of modifying physicochemical and biochemical reactions in the mitochondrial matrix, the mitochondria can be provided with a substrate that increases energy production efficiency and the amount of ATP. Physicochemical parameters of oleic MFA has become the standard of an oxidation substrate for in vivo energy production; this MFA was synthesized in organisms for millions of years. Environmental influences are the second factor which determines kinetic perfection of biological organisms during phylogenesis. Are these influences always beneficial? Mostly, they are not. However, they largely stimulate adaptive functions of the organism, including the biological function of locomotion, cognitive function and the function of positioning in the environment. Biological, energy and kinetic perfection formed in vivo can be easily destroyed if phylogenetically herbivorous Homo sapiens abuses the diet of carnivorous animals (meat) which was not consumed by him and his ancestors during phylogenesis. This abuse is the major cause of metabolic pandemias in human population. They are: insulin resistance, atherosclerosis and atheromatosis, obesity and nonalcoholic disease. The most effective measures preventing metabolic pandemias, cardiac heart disease and myocardial infarction are extremely simple. People should remain herbivorous.

Keyword: fatty liver

Heat shock protein 70 promotes lipogenesis in HepG2 cells.

The increasing prevalence of non-alcoholic disease (NAFLD) has followed the international rise in obesity rates. Multiple mechanisms are involved in NAFLD, including endoplasmic reticulum stress and oxidative stress. Heat shock protein 70 (HSP70), which is abundant in most organisms, is sensitive to stress. However, the role of HSP70 in NAFLD has not been investigated. Here, we investigated the possible role of HSP70 in lipid synthesis.C57BL/6 mice were fed a high-fat diet, and HepG2 cells were treated with 0.5\xa0mM (PA). HSP70 expression was detected by qPCR, Western blot and immunohistochemistry. Total cholesterol (TC) and triglyceride (TG) levels were detected by enzyme-linked immunosorbent assay (ELISA). After Hsp70 overexpression and knockdown, TC and TG levels and FAS, SCD, and ACC expression were detected.HSP70 expression was significantly increased in the livers of obese mice. In vitro, HSP70 expression was markedly induced by PA in HepG2 cells. Notably, HSP70 overexpression in HepG2 cells enhanced TC and TG synthesis, in parallel with the upregulation of lipogenic genes, including FAS, SCD and ACC. By contrast, HSP70 knockdown decreased the levels of cellular lipids and the expression of FAS, SCD, and ACC in HepG2 cells. Together, our results suggest that HSP70 may promote lipogenesis in HepG2 cells.Heat shock protein 70 promotes lipogenesis in HepG2 cells.

Keyword: fatty liver

Changes of the Profile in Erythrocyte Membranes of Patients following 6-Month Dietary Intervention Aimed at the Regression of Nonalcoholic Disease (NAFLD).

Nonalcoholic disease (NAFLD) is closely related to the metabolism disorders of acids. The pathogenesis of the disease includes an increased concentration of FFA in blood, an increase in the biosynthesis of acids, and disorders in the process of -oxidation.The aim of the study was to analyze the acids in erythrocyte membranes among 55 patients with NAFLD who were subjected to a 6-month dietary intervention in order to reduce .Basic anthropometric and biochemical measurements were performed. The profile of acids was measured in the membranes of erythrocytes and analyzed by gas chromatography. The dietary compliance was evaluated using 72-diary questionnaires, anthropometric measurements.With the reduction of (p<0.01), the patients\' biochemical and anthropometric parameters were significantly improved. A significant decrease in the concentration of alanine aminotransferase (p<0.01) and asparagine aminotransferase (p<0.01) was observed, along with a decrease in the amount of insulin (p<0.05) and insulin resistance (p<0.05). Significant changes in terms of the profile were observed among patients who followed the dietary intervention. There was a noticeable tendency in terms of the reduction (p<0.055) and a significant reduction of stearic (p<0.05). Significant changes in the profile of acids were also associated with the reductionof palmitoleic (p<0.05) and oleic acids (p<0.05). Another statistically significant change observed was the increase in polyunsaturated acids. In particular (p<0.01) the rise of eicosapentaenoic (p<0.055) and docosahexaenoic acids (p<0.55) was noted.The profile of acids turned out to be a potential biomarker of the changes during NAFLD regression. Further research is needed to fully elucidate the usefulness and applicability of our findings in the management of NAFLD.

Keyword: fatty liver

Nicotinamide riboside regulates inflammation and mitochondrial markers in AML12 hepatocytes.

The NAD precursor nicotinamide riboside (NR) is a type of vitamin B found in cow\'s milk and yeast-containing food products such as beer. Recent studies suggested that NR prevents hearing loss, high-fat diet-induced obesity, Alzheimer\'s disease, and mitochondrial myopathy. The objective of this study was to investigate the effects of NR on inflammation and mitochondrial biogenesis in AML12 mouse hepatocytes.A subset of hepatocytes was treated with (PA; 250 µM) for 48 h to induce hepatocyte steatosis. The hepatocytes were treated with NR (10 µM and 10 mM) for 24 h with and without PA. The cell viability and the levels of sirtuins, inflammatory markers, and mitochondrial markers were analyzed.Cytotoxicity of NR was examined by PrestoBlue assay. Exposure to NR had no effect on cell viability or morphology. Gene expression of sirtuin 1 (Sirt1) and Sirt3 was significantly upregulated by NR in PA-treated hepatocytes. However, Sirt1 activities were increased in hepatocytes treated with low-dose NR. Hepatic pro-inflammatory markers including tumor necrosis factor-alpha and interleukin-6 were decreased in NR-treated cells. NR upregulated anti-inflammatory molecule adiponectin, and, tended to down-regulate hepatokine fetuin-A in PA-treated hepatocytes, suggesting its inverse regulation on these cytokines. NR increased levels of mitochondrial markers including peroxisome proliferator-activated receptor γ coactivator-1α, carnitine palmitoyltransferase 1, uncoupling protein 2, transcription factor A, mitochondrial and mitochondrial DNA in PA-treated hepatocytes.These data demonstrated that NR attenuated hepatic inflammation and increased levels of mitochondrial markers in hepatocytes.

Keyword: fatty liver

SIRT3 promotes lipophagy and chaperon-mediated autophagy to protect hepatocytes against lipotoxicity.

Lipophagy is a lysosomal lipolytic pathway that complements the actions of cytosolic neutral lipases. Chaperon-mediated autophagy (CMA) triggers lipid droplets (LDs) breakdown, to initiate lipolysis via either cytosolic lipases or macroautophagy. SIRT3, a mitochondrial NAD-dependent deacetylase, regulates the acetylation status and activity of many substrates involving in energy metabolism. However, the role of SIRT3 in regulating lipophagy is controversial. The current study showed that SIRT3 expression was decreased and the macroautophagy flux was blocked in the primary hepatocytes from high-fat diet fed mice and P/O ( and oleic mixture) treated AML12 mouse hepatocytes, compared with the corresponding controls. SIRT3 overexpression promoted macroautophagy in LDs from P/O-treated hepatocytes through activating AMP-activated protein kinase (AMPK) and unc-51-like kinase 1, to boost LDs digestion. Gain of SIRT3 expression stimulated the formation of lysosome-associated membrane protein 2A (LAMP-2A)-heat shock cognate 71\u2009kDa protein (HSC70)-perilipin-2 (PLN2) complex, to promote CMA process and reduce the stability of LDs in hepatocytes. Moreover, SIRT3 reduced the expression of stearoyl-CoA desaturase 1, to suppress lipogenesis. In addition, SIRT3 overexpression promoted LDs dispersion on detyrosinated microtubules, and directly deacetylated long-chain acyl-CoA dehydrogenase to enhance mitochondrial energetics. Taken together, SIRT3 ameliorates lipotoxicity in hepatocytes, which might be a potential target for the treatment of nonalcoholic disease.

Keyword: fatty liver

Transport of Ca and Ca-Dependent Permeability Transition in Rat Mitochondria under the Streptozotocin-Induced Type I Diabetes.

Although diabetes mellitus is known to be a disease associated with mitochondrial dysfunction, not everything is clear about mitochondrial Ca transport and Ca-induced permeability transition in diabetic cells. The objective of this work was to study the operation of MCU and Ca-dependent mitochondrial permeabilization in the cells of Sprague-Dawley rats under the streptozotocin-induced type I diabetes. It was shown that two weeks after the induction of diabetes, the rate of Ca uptake by the mitochondria of diabetic animals increased ~1.4-fold. The expression of MCU and MICU1 subunits did not change, yet the quantity of dominant-negative MCUb channel subunits was almost twice as lower. The organelles also became more resistant to the induction of CsA-sensitive MPT pore and less resistant to the induction of CsA-insensitive palmitate/Ca-induced pore. The mitochondria of diabetic cells also showed changes in the lipid matrix of their membranes. The content of acids in the membranes grew, and microviscosity of the lipid bilayer (assessed with laurdan) increased. At the same time, lipid peroxidation (assessed by the production of malonic dialdehyde) was stimulated. The paper discusses the consequences of the diabetes-related changes in mitochondria in the context of cell physiology.

Keyword: fatty liver

[Effect of lipid-induced macrophage M1/M2 polarization on lipid metabolism in hepatocytes].

This study aims to explore the effect of lipid-induced macrophage M1/M2 polarization on lipid metabolism in hepatocytes. RAW264.7 macrophages were incubated with different kinds of acids including saturated acids- (PA), monounsaturated acids-oleic (OA) and polyunsaturated acids-docosahexaenoic (DHA), and cell culture supernatants were collected to prepare conditioned medium (CM). Hepatocytes were isolated by in situ perfusion of the with collagenase in mice, and a macrophage-hepatocyte CM co-culture system was established. Macrophage M1/M2 phenotype markers were detected by Real-time PCR. Lipid synthesis and decomposition related mRNA and protein expressions in hepatocytes were detected by Real-time PCR and Western Blot. Lipid depositions in hepatocytes were detected by oil red O staining. An analysis of variance was used for comparison of means between multiple groups. Compared with control groups, PA polarized macrophages to a M1 phenotype (expression of TNF-α and IL-6 significantly increased, F≥22.68, P < 0.01), OA polarized macrophages to a M1/M2 mixed phenotype (expression of IL-6, Mrc2 and IL-10 increased F≥4.94, P < 0.05) and DHA polarized macrophages to a M2 phenotype (expression of Mrc2 and IL-10 significantly increased, F≥4.94, P < 0.01). CM-PA significantly increased lipid synthesis related genes, including SREBP1C, ACC1 mRNA expression (F≥5.66, P < 0.01) and FASN, ACC1 protein expression (F≥38.34, P < 0.05) in hepatocytes, and decreased lipid decomposition gene ACOX1 protein expression (F=154.48, P < 0.01). CM-OA affected several lipid metabolism genes expression. CM-DHA significantly increased CPT1A mRNA expression (F = 10.30, P < 0.01) and ACOX1, CPT1A protein expression (F≥47.06, P < 0.05), and decreased SREBP1C, ACC1 protein expression (F≥65.84, P < 0.05) in hepatocytes. Massive lipid droplets were deposited in hepatocytes in CM-PA treated hepatocytes, and a few amount of lipid droplets were deposited in CM-DHA treated hepatocytes. Different acids affect the balance of lipid metabolism in hepatocytes and by inducing macrophage M1 / M2 polarization, thus promoting or delaying the progression of non-alcoholic disease.

Keyword: fatty liver

Upregulation of miR-181a impairs lipid metabolism by targeting PPARα expression in nonalcoholic disease.

Recent studies have reported elevated expression of miR-181a in patients with non-alcoholic disease (NAFLD), suggesting that it may play an important role in lipid metabolism and insulin resistance. We aimed to investigate the effect of miR-181a in lipid metabolism and find new treatments for NAFLD. The expression level of miR-181a in NAFLD patient serum and a (PA)-induced NAFLD cell model was examined by Q-PCR. Oil red O staining and triglyceride assays were used to assess lipid accumulation in hepatocytes. Western blotting was used to detect the protein expression levels of peroxisome proliferator-activated receptor-α (PPARα) and the β-oxidation-related genes. Direct interactions were validated by dual-luciferase reporter gene assays. MiR-181a expression was significantly upregulated in the serum of NAFLD patients and PA-induced hepatocytes. Inhibition of miR-181a expression resulted in the increased expression of PPARα and its downstream genes, and PA-induced lipid accumulation in hepatocytes was also inhibited. Upregulation of miR-181a resulted in the downregulation of its direct target PPARα and downstream gene expression of PPARα as well as aggravated lipid accumulation in hepatocytes. At the same time, the increased expression of PPARα can offset lipid accumulation in hepatocytes induced by miR-181a mimics. This study demonstrates that reducing the expression of miR-181a may improve lipid metabolism in NAFLD. The downregulation of miR-181a expression can be a therapeutic strategy for NAFLD by modulating its target PPARα.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Effects of saturated and omega-3 polyunsaturated acids on Sertoli cell apoptosis.

Obesity is believed to negatively affect male semen quality and is accompanied by dysregulation of free (FFA) metabolism in plasma. However, the implication of dysregulated FFA on semen quality and the involvement of Sertoli cells remain unclear. In the present study, we report obesity decreased Sertoli cell viability through dysregulated FFAs. We observed an increased rate of apoptosis in Sertoli cells, accompanied with elevated FFA levels, in the testes of obese mice that were provided a high-fat diet (HFD). Moreover, the levels of reactive oxygen species were elevated. Furthermore, we demonstrated by in vitro assays that saturated (PA), which is the most common saturated FFA in plasma, led to decreased cell viability of TM4 Sertoli cells in a time- and dose-dependent manner. A similar finding was noted in primary mouse Sertoli cells. In contrast to saturated FFA, omega-3 (ω-3) polyunsaturated acids (PUFAs) protected Sertoli cells from PA-induced lipotoxicity at the physiologically relevant levels. These results indicated that the lipotoxicity of saturated acids might be the cause of obesity-induced Sertoli cell apoptosis, which leads to decreased semen quality. In addition, ω-3 PUFAs could be classified as protective FFAs.FFA: free ; HFD: high-fat diet; SD: standard diet; PA: ; PUFA: polyunsaturated ; AI: apoptotic index; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide; ROS: reactive oxygen species; HE: Hematoxylin and eosin; WT1: Wilm Tumor 1; NAFLD: non- alcoholic disease; DCFH-DA: 2\', 7\' dichlorofluorescin diacetate; 36B4: acidic ribosomal phosphoprotein P0; SD: standard deviation; EPA: eicosapentaenoic ; PI: propidium iodide; DHA: docosahexenoic .

Keyword: fatty liver

activates NLRP3 inflammasomes in mouse Kupffer cells through mitochondrial DNA release.

Nonalcoholic disease (NAFLD) is the most common disease in many developed and developing countries worldwide. It has been well established that the chronic sterile inflammation caused by the NLRP3 inflammasome is closely related to NAFLD development. Kupffer cells (KCs) are involved in the pathogenesis of various diseases. We used methionine choline-deficient diets to establish a mouse nonalcoholic steatohepatitis (NASH) model. The expression and formation of the NLRP3 inflammasome in the KCs from the mouse and cell models were determined by Western blotting and co-immunoprecipitation. Evidence of mitochondrial DNA (mtDNA) release was determined by live cell labeling and imaging. KCs and the NLRP3 inflammasome exerted proinflammatory effects on the development and progression of NASH through secretion of the proinflammatory cytokine IL-1β. NLRP3, ASC and Caspase-1 protein expression levels in KCs from NASH mouse livers were significantly higher than those in KCs from NLRP3 mice, and the number of NLRP3 inflammasome protein complexes was significantly higher in KCs from NASH mouse livers, whereas these protein complexes could not be formed in NLRP3 mice. In in vitro experiments, (PA) decreased the mitochondrial membrane potential and subsequently induced mtDNA release from the mitochondria to the cytoplasm. NLRP3 inflammasome expression was substantially increased, and mtDNA-NLRP3 inflammasome complexes formed upon PA stimulation. Our data suggest that mtDNA released from mitochondria during PA stimulation causes NLRP3 inflammasome activation, providing a missing link between NLRP3 inflammasome activation and NASH development, via binding of cytosolic mtDNA to the NLRP3 inflammasome.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Pro-Inflammatory CXCR3 Impairs Mitochondrial Function in Experimental Non-Alcoholic Steatohepatitis.

Mitochondrial dysfunction plays a crucial role in the development of non-alcoholic steatohepatitis (NASH). However, the regulator of mitochondrial dysfunction in the pathogenesis of NASH is still largely unclear. CXCR3 is an essential pro-inflammatory factor in chronic diseases. We explored the significance of CXCR3 in regulating mitochondrial function during NASH development in animal models and cultured hepatocytes.The effects of CXCR3 on mitochondrial function were evaluated by genetic knockout or pharmacological inhibition in mouse models and . The ultrastructural changes of mitochondria were assessed by transmission electron microscopy (TEM). Hepatic levels of mitochondrial reactive oxygen species (ROS), DNA damage, membrane potential and ATP were examined.CXCR3 ablation by genetic knockout or pharmacological inhibition in mice protected against NASH development by influencing mitochondrial function. Similarly, depletion of CXCR3 reduced steatohepatitis injury in cultured hepatocytes. TEM analysis revealed that mitochondrial integrity was much improved in CXCR3 knockout (CXCR3) compared to wildtype (WT) mice. In agreement with this, impaired mitochondrial function was pronounced in WT mice compared to CXCR3 mice, evidenced by increased protein expression of dynamic-related protein-1 (DRP1) and fission-1 (FIS1) and decreased protein expression of mitofusin-1 (MFN1). Mitochondrial dysfunction was induced in AML-12 hepatocytes by methionine and choline deficient medium and in HepG2 cells by . The impaired mitochondrial function in both cell lines was evidenced by reduced membrane potential and ATP content, and by increased mitochondrial ROS accumulation and DNA damage. However, CXCR3 knockdown by siCXCR3 significantly diminished the mitochondrial dysfunction in both AML-12 and HepG2 hepatocytes. In addition, inhibition of CXCR3 by CXCR3 specific antagonists SCH546738 and AMG487 restored mitochondrial function and inhibited mitochondrial-dependent apoptosis in the of WT mice fed with methionine and choline deficient diet.CXCR3 induces mitochondrial dysfunction, which contributes to the pathogenesis of steatohepatitis. Pharmacologic blockade of CXCR3 prevents mitochondrial dysfunction and restores the severity of steatohepatitis, indicating a potential clinical impact for controlling the disease.

Keyword: fatty liver

Mst1 inhibition attenuates non-alcoholic disease via reversing Parkin-related mitophagy.

Obesity-related non-alcoholic disease (NAFLD) is connected with mitochondrial stress and hepatocyte apoptosis. Parkin-related mitophagy sustains mitochondrial homeostasis and hepatocyte viability. However, the contribution and regulatory mechanisms of Parkin-related mitophagy in NAFLD are incompletely understood. Macrophage stimulating 1 (Mst1) is a novel mitophagy upstream regulator which excerbates heart and cancer apoptosisn via repressing mitophagy activity. The aim of our study is to explore whether Mst1 contributes to NAFLD via disrupting Parkin-related mitophagy. A NAFLD model was generated in wild-type (WT) mice and Mst1 knockout (Mst1-KO) mice using high-fat diet (HFD). Cell experiments were conducted via (PA) treatment in the primary hepatocytes. The results in our study demonstrated that Mst1 was significantly upregulated in HFD-treated livers. Genetic ablation of Mst1 attenuated HFD-mediated hepatic injury and sustained hepatocyte viability. Functional studies illustrated that Mst1 knockdown reversed Parkin-related mitophagy and the latter protected mitochondria and hepatocytes against HFD challenge. Besides, we further figured out that Mst1 modulated Parkin expression via the AMPK pathway; blockade of AMPK repressed Parkin-related mitophagy and recalled hepatocytes mitochondrial apoptosis. Altogether, our data identified that NAFLD was closely associated with the defective Parkin-related mitophagy due to Mst1 upregulation. This finding may pave the road to new therapeutic modalities for the treatment of disease.Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

Keyword: fatty liver

γδ T cells Promote Steatohepatitis by Orchestrating Innate and Adaptive Immune Programming.

The recruitment and activation of inflammatory cells in the delineates the transition from hepatic steatosis to steatohepatitis. We found that in steatohepatitis, γδT cells are recruited to the by CCR2, CCR5, and NOD2 signaling and are skewed towards an IL-17A phenotype in an ICOS-ICOSL dependent manner. γδT cells exhibit a distinct Vγ4 , PD1 , Ly6C CD44 phenotype in steatohepatitis. Moreover, γδT cells upregulate both CD1d, which is necessary for lipid-based antigens presentation, and the free receptor CD36. γδT cells are stimulated to express IL-17A by and CD1d ligation. Deletion, depletion, and targeted interruption of γδT cell recruitment protects against diet-induced steatohepatitis and accelerates disease resolution. We demonstrate that hepatic γδT cells exacerbate steatohepatitis, independent of IL-17 expression, by mitigating conventional CD4 T cell expansion and modulating their inflammatory program via CD1d-dependent VEGF expression.© 2019 by the American Association for the Study of Diseases.

Keyword: fatty liver

Hsp72 protects against injury via attenuation of hepatocellular death, oxidative stress, and JNK signaling.

Heat shock protein (Hsp) 72 is a molecular chaperone that has broad cytoprotective functions and is upregulated in response to stress. To determine its hepatic functions, we studied its expression in human disorders and its biological significance in newly generated transgenic animals.Double transgenic mice overexpressing Hsp72 (gene Hspa1a) under the control of a tissue-specific tetracycline-inducible system (Hsp72-LAP mice) were produced. Acute injury was induced by a single injection of acetaminophen (APAP). Feeding with either a methionine choline-deficient (MCD; 8\u202fweeks) or a 3,5-diethoxycarbonyl-1,4-dihydrocollidine-supplemented diet (DDC; 12\u202fweeks) was used to induce lipotoxic injury and Mallory-Denk body (MDB) formation, respectively. Primary hepatocytes were treated with .Patients with non-alcoholic steatohepatitis and chronic hepatitis C infection displayed elevated HSP72 levels. These levels increased with the extent of hepatic inflammation and HSP72 expression was induced after treatment with either interleukin (IL)-1β or IL-6. Hsp72-LAP mice exhibited robust, hepatocyte-specific Hsp72 overexpression. Primary hepatocytes from these animals were more resistant to isolation-induced stress and Hsp72-LAP mice displayed lower levels of hepatic injury in vivo. Mice overexpressing Hsp72 had fewer APAP protein adducts and were protected from oxidative stress and APAP-/MCD-induced cell death. Hsp72-LAP mice and/or hepatocytes displayed significantly attenuated Jnk activation. Overexpression of Hsp72 did not affect steatosis or the extent of MDB formation.Our results demonstrate that HSP72 induction occurs in human disease, thus, HSP72 represents an attractive therapeutic target owing to its broad hepatoprotective functions.HSP72 constitutes a stress-inducible, protective protein. Our data demonstrate that it is upregulated in patients with chronic hepatitis C and non-alcoholic steatohepatitis. Moreover, Hsp72-overexpressing mice are protected from various forms of stress.Copyright © 2018 European Association for the Study of the . All rights reserved.

Keyword: fatty liver

High Glucose Concentration Impairs 5-PAHSA Activity by Inhibiting AMP-Activated Protein Kinase Activation and Promoting Nuclear Factor-Kappa-B-Mediated Inflammation.

Recently, the endogenous -5-hydroxystearic (5-PAHSA) was found to increase insulin sensitivity and have anti-inflammatory effects in mice with high-fat diet (HFD)-induced diabetes. However, it is unknown if 5-PAHSA affects glucose and lipid metabolism in db/db mice, which are characterized by extreme hyperglycemia. Here, we aim to determine the effect of continued 5-PAHSA administration on glucose and lipid metabolism in db/db mice. We also used 3T3-L1 cells and HepG2 cells to investigate the mechanism behind this effect. HepG2 cells and 3T3-L1 cells were induced to become models of insulin resistance. The models were used to test the effect of 5-PAHSA on insulin signaling. 5-PAHSA was administered orally to db/db mice for 1 month to assess its effects on glucose and lipid metabolism. We also exposed HepG2 cells to high glucose concentrations to investigate the influence on 5-PAHSA\'s effects on hepatic lipid metabolism and inflammation. 5-PAHSA improved glucose uptake and insulin signaling in HepG2 cells and 3T3-L1 cells. However, after 1 month of treatment, 5-PAHSA did not reduce blood glucose levels, but increased inflammation and promoted in db/db mice. In HepG2 cells under normal glucose conditions, 5-PAHSA treatment reduced lipogenesis and increased lipid oxidation. Notably, a high glucose concentration in cell media abolished the positive effects of 5-PAHSA treatment. These changes were associated with: decreased phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC); upregulation of sterol-regulatory element-binding protein 1c (SREBP1c), and synthase (FAS); and downregulation of carnitine palmitoyltransferase 1 (CPT1). Besides, the anti-inflammatory effect of 5-PAHSA was also impaired by high glucose conditions. Thus, high glucose concentrations impaired 5-PAHSA action by inhibiting the AMPK signaling pathway and promoting nuclear factor-kappa-B (NF-κB) mediated inflammation.

Keyword: fatty liver

Omega-3 polyunsaturated acids alleviate hepatic steatosis-induced inflammation through Sirt1-mediated nuclear translocation of NF-κB p65 subunit in hepatocytes of large yellow croaker (Larmichthys crocea).

Hepatic steatosis induced inflammation is becoming increasingly prevalent in farmed fish. This study was conducted to investigate the protective effects of omega-3 polyunsaturated acids (ω-3 PUFAs) against hepatic steatosis-induced inflammation and its potential molecular mechanisms in hepatocyte of large yellow croaker (Larmichthys crocea). We found that the hepatic steatosis-induced inflammation was relieved by ω-3 PUFAs, meanwhile, the Sirt1 activity and transcript expression was increased by ω-3 PUFAs. The increased Sirt1 activity can decrease the hepatic steatosis-induced inflammation. The protective effects of ω-3 PUFAs against hepatic steatosis-induced inflammation was reversed by the treatment with Sirt1 inhibitor EX-527. The nuclear translocation of nuclear transcription factor kappa-B (NF-κB) p65 was significantly decreased after ω-3 PUFAs treatments compared to the stimulation group. The ω-3 PUFAs induced cytoplasm translocation of NF-κB p65 was reversed by EX-527. Together, ω-3 PUFAs alleviate hepatic steatosis-induced inflammation through Sirt1-mediated nuclear translocation of NF-κB p65 subunit in hepatocytes of large yellow croaker. The present study provides important insight into the mechanisms of the protective effects of ω-3 PUFAs, providing theory bases for alleviating the hepatic steatosis induced inflammation of farmed fish, thereby offering great benefits to the aquaculture industry and fish consumers.Copyright © 2017. Published by Elsevier Ltd.

Keyword: fatty liver

Long-chain activates hepatocytes through CD36 mediated oxidative stress.

Accumulating evidence suggests that activated hepatocytes are involved in the deposition of the excess extracellular matrix during fibrosis via the epithelial to mesenchymal transition. Lipid accumulation in hepatocytes are implicated in the pathogenesis of chronic injury. CD36 is known to mediate long-chain (LCFA) uptake and lipid metabolism. However, it is unclear whether LCFA directly promotes hepatocyte activation and the involved mechanisms have not been fully clarified.Mice were fed with a high fat diet (HFD) and normal hepatocyte cells (Chang cells) were treated with (PA) in vivo and in vitro. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to examine the gene and protein expression of molecules involved in hepatic fibrogenesis and hepatocyte activation. CD36 was knocked down by transfecting CD36 siRNA into hepatocyte cells. Hydrogen peroxide (HO) and reactive oxygen species (ROS) levels were detected using commercial kits.HFD induced a profibrogenic response and up-regulated CD36 expression in vivo. Analogously, PA increased lipid accumulation and induced human hepatocyte activation in vitro, which was also accompanied by increased CD36 expression. Interestingly, knockdown of CD36 resulted in a reduction of hepatocyte lipid deposition and decreased expression of Acta2 (34% decrease), Vimentin (29% decrease), Desmin (60% decrease), and TGF-β signaling pathway related genes. In addition, HFD and PA increased the production of HO in vivo (48% increase) and in vitro (385% increase), and the antioxidant, NAC, ameliorated PA-induced hepatocyte activation. Furthermore, silencing of CD36 in vitro markedly attenuated PA-induced oxidative stress (HO: 41% decrease; ROS: 39% decrease), and the anti-activation effects of CD36 knockdown could be abolished by pretreatment with HO.Our study demonstrated that LCFA facilitates hepatocyte activation by up-regulating oxidative stress through CD36, which could be an important mechanism in the development of hepatic fibrosis.

Keyword: fatty liver

Adaptations of hepatic lipid metabolism and mitochondria in dairy cows with mild .

The inevitable deficiency in nutrients and energy at the onset of lactation requires an optimal adaptation of the hepatic metabolism to overcome metabolic stress. is one of the main health disorders after parturition. Therefore, to investigate changes in hepatic lipid metabolic status and mitochondria in dairy cows with mild , and blood samples were collected from healthy cows (n = 15) and cows with mild (n = 15). To determine the effects of acids (PA), one of the major component of acids, on lipid metabolism and mitochondria in vitro, calf hepatocytes were isolated from healthy calves and treated with various concentrations of PA (0, 50, 100, and 200 μM). Dairy cows with mild displayed hepatic lipid accumulation. The protein levels of sterol regulatory element-binding protein 1c (SREBP-1c) and peroxisome proliferator-activated receptor-α (PPARα) and mRNA levels of acetyl CoA carboxylase 1 (ACC1), synthase (FAS), acyl-CoA oxidase (ACO), and carnitine palmitoyltransferase 1A (CPT1A) were significantly higher in dairy cows with mild than in control cows. The hepatic mitochondrial DNA content, mRNA levels of oxidative phosphorylation complexes I to V (CO 1-V), protein levels of cytochrome c oxidase subunit IV (COX IV), voltage dependent anion channel 1 (VDAC1), peroxisome proliferator activated receptor-γ coactivator-1α (PGC-1α) and nuclear respiratory factor 1 (NRF1), and adenosine triphosphate (ATP) content were all markedly increased in the of dairy cows with mild compared with healthy cows. The PA treatment significantly increased lipid accumulation; protein levels of SREBP-1c and PPARα; and mRNA levels of ACC1, FAS, ACO, and CPT1A in calf hepatocytes. Moreover, the mitochondrial DNA content, mRNA levels of CO 1-V, protein levels of COX IV, VDAC1, PGC-1α, NRF1, mitochondrial transcription factor A, and ATP content were significantly increased in PA-treated hepatocytes compared with control hepatocytes. The protein level of mitofusin-2 was significantly decreased in PA-treated groups. In conclusion, lipid synthesis and oxidation, number of mitochondria, and ATP production were increased in the of dairy cows with mild and PA-treated calf hepatocytes. These changes in hepatic mitochondria and lipid metabolism may be the adaptive mechanism of dairy cows with mild .Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fatty liver

MicroRNA-194 inhibition improves dietary-induced non-alcoholic disease in mice through targeting on FXR.

Non-alcoholic disease (NAFLD) affects obesity-associated metabolic syndrome, which exhibits hepatic steatosis, insulin insensitivity and glucose intolerance. Previous studies indicated that hepatic microRNAs (miRs) play critical roles in the development of NAFLD. In this study, we aim to explore the pathophysiological role of miR-194 in obesity-mediated metabolic dysfunction. Our findings show that the high fat diet or treatment significantly increase hepatic miR-194 levels in vivo and in vitro. Silence of miR-194 protects -induced inflammatory response in cultured hepatocytes, and attenuates structural disorders, lipid deposits and inflammatory response in . MiR-194 inhibitor also improves glucose and insulin intolerance in obese mice. Through dual luciferase assay, we demonstrate that miR-194 directly binds to FXR/Nr1h4 3\'-UTR, and inhibits gene expression of FXR/Nr1h4. Furthermore, overexpression of miR-194 downregulates FXR/Nr1h4 in cultured hepatocytes, but miR-194 inhibitor reversely increases FXR/Nr1h4 expression in obese mouse tissues. On the contrast, silence of FXR/Nr1h4 abolishes the hepatic benefits in obese mice treated with miR-194 inhibitor. Present study provides a novel finding that suppression of miR-194 attenuates dietary-induced NAFLD via upregulation of FXR/Nr1h4. The findings suggest miR-194/FXR are potential diagnostic markers and therapeutic targets for NAFLD.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Resveratrol Ameliorates Lipid Droplet Accumulation in Through a SIRT1/ ATF6-Dependent Mechanism.

Lipid droplets (LDs) are dynamic organelles that store neutral lipids during times of energy excess, and an increased accumulation of LDs in the is closely linked to hepatic steatosis. Our previous studies suggested that resveratrol (RSV) supplement could improve hepatic steatosis, but the underlying mechanism, particularly which related to LD accumulation, has not yet been elucidated.A high-fat diet (HFD) and were used to induce hepatic steatosis in mouse and hepatocytes, respectively. The effects of RSV on LD accumulation were analyzed in vivo and in vitro. The effects of RSV on the expression levels of LD-associated genes (ATF6, Fsp27β/CIDEC, CREBH, and PLIN1) were measured by qRT-PCR and western blot assays, followed by KD or overexpression of SIRT1 and ATF6 with small interfering RNAs or overexpressed plasmids, respectively. The dual luciferase reporter assay, chromatin immunoprecipitation assay, coimmunoprecipitation, and proximity ligation assay were utilized to clarify the mechanism of transcriptional regulation and possible interaction between SIRT1 and ATF6.There was a significant increase in the accumulation of LDs in and hepatocytes during the process of HFD-induced steatosis, respectively, which was significantly inhibited by RSV supplementation. RSV notably activated SIRT1 expression and decreased the expression levels of ATF6, Fsp27β/CIDEC, CREBH, and PLIN1, which are associated with LD accumulation. Interestingly, the inhibitory effects of RSV on LD accumulation and the associated expression of genes in hepatocytes were abrogated or strengthened with SIRT1 silencing or overexpression, respectively. On the contrary, the benefits of RSV in hepatocytes were eliminated or aggravated when transfected with the overexpressed ATF6 or ATF6 siRNA, respectively. Furthermore, we found that RSV stimulated SIRT1 expression significantly, which was followed by increased deacetylation and inactivation of ATF6, resulting in a positive feedback loop for SIRT1 transcription associated with ATF6 binding to the SIRT1 promoter region.Taken together, these findings indicate that RSV supplementation improves hepatic steatosis by ameliorating the accumulation of LDs, and this might be partially mediated by a SIRT1/ATF6-dependent mechanism.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fatty liver

Insulin-like growth factor binding protein 7 accelerates hepatic steatosis and insulin resistance in non-alcoholic fatty liver disease.

An association between increased insulin-like growth factor binding protein-7 (IGFBP7) expression and insulin resistance in metabolic diseases has been reported. However, the role and molecular mechanism of IGFBP-7 in non-alcoholic fatty liver disease (NAFLD) remains largely unknown. Therefore, the potential function of IGFBP7 in the pathological progression of NAFLD was explored in this investigation. For in vivo experiments, an animal model of NAFLD was established in C57BL/6 mice by feeding a high-fat diet (HFD), and IGFBP7 was knocked down by injecting adeno-associated adenovirus (AAV)-mediated short-hairpin (sh)-IGFBP7 into the liver. We found that AAV-sh-IGFBP7 treatment significantly alleviated hepatocyte injury and inhibited hepatic lipid accumulation by reducing lipogenesis-associated gene expression. Furthermore, downregulation of IGFBP7 markedly ameliorated IR and restored impaired insulin signalling by elevating the phosphorylation levels of IRS-1, Akt and GSK3β in HFD-treated mice. Similar results were also confirmed by an in vitro study in a (PA)-stimulated HepG2 cell model. In conclusion, our study demonstrates that IGFBP7 contributes to hepatic steatosis and insulin resistance in NAFLD development, which might serve as a novel therapeutic agent for the treatment of NAFLD.© 2019 John Wiley & Sons Australia, Ltd.

Keyword: fatty liver

Systematic evaluation of phenolic compounds and protective capacity of a new mulberry cultivar J33 against -induced lipotoxicity using a simulated digestion method.

This research aimed to investigate the protective effects of a new mulberry cultivar J33 with simulated gastrointestinal digestion against (PA)-induced lipotoxicity. LC-MS analysis revealed that the contents of four flavonoid glycosides (quercetin rhamnosylhexoside hexoside, quercetin rhamnosylhexoside, quercetin hexoside, kaempferol rhamnosylhexoside) increased after digestion. Besides, mulberry digest (MBD) at 0.5-2\u202fmg/mL significantly reduced PA-induced lipotoxicity in human hepatocytes, while mulberry extract without digestion (MBE) showed no protection. Further investigations demonstrated that the protection of MBD was attributed to two aspects. On the one hand, MBD could attenuate PA-induced oxidative stress by suppressing ROS accumulation, regulating intracellular glutathione and ameliorating mitochondrial dysfunction. On the other hand, MBD could promote PA incorporation into inert triglycerides (TG) to deal with the acute lipid overload, reducing the lipotoxicity caused by PA. Overall, our research might provide a new perspective of mulberry cultivar J33 in ameliorating non-alcoholic disease (NAFLD).Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

PKCδ silencing alleviates saturated induced ER stress by enhancing SERCA activity.

Protein kinase C δ (PKCδ) plays an important role in nonalcoholic disease (NAFLD), however, the mechanism remains unknown. The present study explored the role of PKCδ in NAFLD development and investigated the relationships between PKCδ, calcium homeostasis, and endoplasmic reticulum (ER) stress (ERS). Hepatic steatosis cell model was induced by (PA) in L02 cells. Lipid accretion was evaluated using Oil Red O staining and a triglyceride (TG) detection kit. PKCδ was down-regulated by siRNA. RT-PCR and Western blotting were used to detect the expression of ERS markers. The fluorescence of Ca influx was recorded using confocal microscopy. Sarco-ER Ca-ATPase (SERCA) activity was measured by ultramicro-ATP enzyme test kit. PA treatment induced lipid accretion in L02 cells, destroyed the ER structure, and increased PKCδ activation in a time-dependent manner. Further, PA treatment significantly increased the expression of ERS markers, Ig heavy chain binding protein (Bip), and homologous proteins of CCAAT-enhancer binding proteins (CHOP). PKCδ silencing down-regulated Bip and CHOP expression, indicating a successful alleviation of ERS. The increased calcium storage induced by PA stimulation was significantly decreased in L02 cells treated with PKCδ siRNA compared with the negative control. Moreover, diminished SERCA activity caused by PA was recovered in PKCδ siRNA transfected cells. To the best of our knowledge, this is the first report demonstrating that the inhibition of PKCδ alleviates ERS by enhancing SERCA activity and stabilizing calcium homeostasis.© 2017 The Author(s).

Keyword: fatty liver

The Hepatotoxicity of in Zebrafish Involves the Intestinal Microbiota.

(PA) is the main saturated naturally occurring in animal fats and vegetable oils. In recent decades, palm oil, an alternative lipid source containing high amounts of PA, has been widely used to replace fish oil in aquafeed.We investigated the hepatotoxicity of PA in zebrafish and the underlying mechanism.One-month-old zebrafish fed a high-fat diet (HFD) containing 16% soybean oil and 3 PA-incorporated HFDs [4%, 8%, and 12% PA (12PA)] for 2 wk (experiment 1) and 4 wk (experiment 2) were used to evaluate PA-induced damage and endoplasmic reticulum (ER) stress. Germ-free (GF) zebrafish fed low-fat, high-fat, or 12PA diets for 5 d were used to study the direct effects of PA on damage (experiment 3). GF zebrafish colonized with HFD or 12PA microbiota for 48 h were used to elucidate the indirect effects of PA-altered microbiota on damage (experiment 4). Last, GF zebrafish colonized with HFD or 12PA microbiota were used to evaluate the effects of different microbiotas on PA absorption (experiment 5).In experiment 1, the proportion of PA in the linearly increased as its percentage in dietary lipid increased (r2\xa0=\xa00.83, P\xa0<\xa00.05). In experiment 2, the expression of glucose-regulated protein 78 (Grp78) and C/EBP-homologous protein (Chop) was higher in the 12PA group than in the HFD group (2.2- and 2.7-fold, respectively; P\xa0<\xa00.05). The activity of caspase-12 was increased by 61.1% in the 12PA group compared with the HFD group (P\xa0<\xa00.05). In experiment 3, caspase-12 activity was higher in the 12PA group than in the HFD group (P\xa0<\xa00.05). In experiment 4, GF zebrafish colonized with PA-altered microbiota had higher caspase-12 activity (P\xa0<\xa00.05) than those colonized by HFD microbiota. In experiment 5, PA-altered microbiota promoted PA absorption (P\xa0<\xa00.05) and aggravated ER stress and damage in the context of high-PA feeding.The PA-altered microbiota indirectly induced ER stress and damage in zebrafish. Moreover, the PA microbiota promoted the absorption of PA, leading to enhanced PA overflow into the and aggravated hepatotoxicity of PA in zebrafish.

Keyword: fatty liver

Analysis of proautophagic activities of Citrus flavonoids in cells reveals the superiority of a natural polyphenol mixture over pure flavones.

Autophagy dysfunction has been implicated in the pathogenesis of nonalcoholic disease (NAFLD). Natural compounds present in bergamot polyphenol fraction (BPF) prevent NAFLD and induce autophagy in rat livers. Here, we employed HepG2 cells expressing DsRed-LC3-GFP, a highly sensitive model system to screen for proautophagic compounds present in BPF. BPF induced autophagy in a time- and dose-dependent fashion and the effect was amplified in cells loaded with . Autophagy was mediated by the hydrophobic fraction of -hydrolyzed BPF (A-BPF), containing six flavanone and flavone aglycones as identified by liquid chromatography-high-resolution mass spectrometry. Among them, naringenin, hesperitin, eriodictyol and diosmetin were weak inducers of autophagy. Apigenin showed the strongest and dose-dependent proautophagic activity at early time points (6 h). Luteolin induced a biphasic autophagic response, strong at low doses and inhibitory at higher doses. Both flavones were toxic in HepG2 cells and in differentiated human progenitors HepaRG upon longer treatments (24 h). In contrast, BPF and A-BPF did not show any toxicity, but induced a persistent increase in autophagic flux. A mixture of six synthetic aglycones mimicking A-BPF was sufficient to induce a similar autophagic response, but it was mildly cytotoxic. Thus, while six main BPF flavonoids fully account for its proautophagic activity, their combined effect is not sufficient to abrogate cytotoxicity of individual compounds. This suggests that a natural polyphenol phytocomplex, such as BPF, is a safer and more effective strategy for the treatment of NAFLD than the use of pure flavonoids.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

The beneficial effects of resveratrol on steatosis and mitochondrial oxidative stress in HepG2 cells.

Nonalcoholic disease (NAFLD) is currently one of the most common chronic diseases, especially in developed countries. One group of substances with a potential use in the treatment of NAFLD are plant polyphenols, represented by resveratrol. The aim of this study was to evaluate the effect of resveratrol on steatosis and oxidative stress in HepG2 cells. The steatosis of cells was carried out using free acids: oleic or and their mixtures. Steatosis was visualized using the intracellular lipid staining by Nile Red dye with a fluorescence microscope. This study also determined the viability of cells and mitochondrial membrane potential. The current study showed that acids and their mixtures induced fat overloading in HepG2 cells. In the group of cells incubated with oleic (OA), observed changes were moderate with prevailing micro-vesicular steatosis. In case of cells incubated with (PA) and the mixtures of acids, micro- and macro-vacuolar steatosis occurred in most of the cells. Resveratrol decreased steatosis in HepG2 cells induced by OA, PA, as well as their mixtures, and in most of experimental groups did not reduce cells viability. Resveratrol reduced the oxidative stress in HepG2 cells treated with acids mixtures.

Keyword: fatty liver

Liraglutide ameliorates non-alcoholic steatohepatitis by inhibiting NLRP3 inflammasome and pyroptosis activation via mitophagy.

Non-alcoholic steatohepatitis (NASH) is a key step in the progression of non-alcoholic disease (NAFLD), which causes serious health problems worldwide. The nucleotide-binding oligomerization domain, leucine-rich repeat-containing receptor-containing pyrin domain 3 (NLRP3) inflammasome and pyroptosis play crucial roles in the progression of NASH. Our team has provided clinical evidence of the effects of glucagon-like peptide-1 (GLP-1) on the improvement in function and histological resolution of NAFLD. Preliminary work has demonstrated that GLP-1 inhibited NLRP3 inflammasome activation in a mouse model of NAFLD. We further explored the potential molecular mechanisms underlying the anti-inflammatory effect of liraglutide, a long-acting GLP-1 analog, in the treatment of NASH. We established a HepG2 cell model of NASH using double stimulation with and lipopolysaccharide to assess NLRP3 inflammasome and pyroptotic cell activity and to evaluate mitochondrial function and mitophagy. Liraglutide reduced lipid accumulation, inhibited NLRP3 inflammasome and pyroptosis activation, attenuated mitochondrial dysfunction and reactive oxygen species generation, augmented mitophagy in hepatocytes. Mitophagy inhibition with 3-methyladenine/PINK1-directed siRNA weakened the liraglutide-mediated suppression of inflammatory injury. We propose that liraglutide suppresses NLRP3 inflammasome-induced hepatocyte pyroptosis via mitophagy to slow the progression of NASH.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Inhibition of hepatocyte nuclear factor 1b induces hepatic steatosis through DPP4/NOX1-mediated regulation of superoxide.

Nonalcoholic disease (NAFLD) is the most common chronic disorder that is closely associated with insulin resistance and type 2 diabetes. Previous studies have suggested that hepatocyte nuclear factor 1b (HNF1b) ameliorates insulin resistance. However, the role of HNF1b in the regulation of lipid metabolism and hepatic steatosis remains poorly understood. We found that HNF1b expression was decreased in steatotic livers. We injected mice with lentivirus (LV) expressing HNF1b shRNA to generate mice with hepatic knockdown of HNF1b. We also injected high fat (HF) diet-induced obese and db/db diabetic mice with LV expressing HNF1b to overexpress HNF1b. Knockdown of HNF1b increased hepatic lipid contents and induced insulin resistance in mice and in hepatocytes. Knockdown of HNF1b worsened HF diet-induced increases in hepatic lipid contents, injury and insulin resistance in mice and PA-induced lipid accumulation and impaired insulin signaling in hepatocytes. Moreover, overexpression of HNF1b alleviated HF diet-induced increases in hepatic lipid content and insulin resistance in mice. Knockdown of HNF1b increased expression of genes associated with lipogenensis and endoplasmic reticulum (ER) stress. DPP4 and NOX1 expression was increased by knockdown of HNF1b and HNF1b directly bound with the promoters of DPP4 and NOX1. Overexpression of DPP4 or NOX1 was associated with an increase in lipid droplets in hepatocytes and decreased expression of DPP4 or NOX1 suppressed the effects of knockdown of HNF1b knockdown on triglyceride (TG) formation and insulin signaling. Knockdown of HNF1b increased superoxide level and decreased glutathione content, which was inhibited by downregulation of DPP4 and NOX1. N-acetylcysteine (NAC) suppressed HNF1b knockdown-induced ER stress, TG formation and insulin resistance. (PA) decreased HNF1b expression which was inhibited by NAC. Taken together, these studies demonstrate that HNF1b plays an essential role in controlling hepatic TG homeostasis and insulin sensitivity by regulating DPP4/NOX1mediated generation of superoxide.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Cangju Qinggan Jiangzhi Decoction Reduces the Development of NonAlcoholic Steatohepatitis and Activation of Kupffer Cells.

Nonalcoholic steatohepatitis (NASH) is defined as lipid accumulation with hepatic injury, inflammation and early to moderate fibrosis. Kupffer cells play a crucial role in promoting hepatic inflammation, which further facilitates the development of NASH. Here we investigated the effects of Cangju Qinggan Jiangzhi decoction (CQJD) on high fat diet (HFD) and methionine-choline deficient (MCD) induced mouse NASH pathogenesis.Mouse NASH models were developed by HFD and MCD diet. The treated mice were divided into three groups: the control group (n = 10), the low-dose CQJD treatment group (n = 10) and the high-dose CQJD treatment group (n = 10). The hepatic injury, inflammation, and apoptotic molecules were evaluated by H&E staining, immunohistochemistry and real-time PCR. Kupffer cells were isolated from control mice and CQJD-treated mice after stimulation by lipopolysaccharide (LPS) and/or . The level of the inflammatory cytokines TNFα, IL1β, and CCL2 was measured by ELISA.The HFD-fed mice displayed significant metabolic, inflammatory, and oxidative stress-related alterations due to hepatic lipid accumulation. CQJD treatment largely normalized the hepatic injury, lowered the ALT/AST level, and reduced the severity of inflammation, as revealed by the decreased inflammatory cytokines levels. In vitro, CQJD blocked the activation of LPS- or -primed Kupffer cells in a dose-dependent manner. In the MCD diet-induced NASH mice, similar therapeutic effects of CQJD were also observed.CQJD ameliorates mouse nonalcoholic steatohepatitis. The reduction in injury and inflammation induced by CQJD is associated with reduced activation of Kupffer cells. Our results suggest that CQJD is a promising therapeutic strategy in clinical steatohepatitis.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fatty liver

Effects of abomasal infusions of acids and one-carbon donors on hepatic ceramide and phosphatidylcholine in lactating Holstein dairy cows.

Our objectives were to (1) determine whether the abomasal infusion of behenic (C22:0) elevated hepatic ceramide relative to (C16:0) or docosahexaenoic (C22:6n-3) infusion; (2) assess whether the abomasal infusion of choline chloride or l-serine elevated hepatic phosphatidylcholine (PC) in cows abomasally infused with C16:0; and (3) characterize the PC lipidome in cows abomasally infused with C22:6n-3, relative to C16:0 or C22:0 infusion. In a 5 × 5 Latin square design, 5 rumen-cannulated Holstein cows (214 ± 4.9 DIM; 3.2 ± 1.1 parity) were enrolled in a study with 6-d periods. Abomasal infusates consisted of (1) (PA; 98% C16:0); (2) PA + choline chloride (PA+C; 50 g/d choline chloride); (3) PA + l-serine (PA+S; 170 g/d l-serine); (4) behenic (BA; 92% C22:0); and (5) an algal oil rich in docosahexaenoic (DHA; 44% C22:6n-3). Emulsion infusates provided 301 g/d of total acids containing a minimum of 40 g/d of C16:0. Cows were fed a corn silage-based diet. Milk was collected on d -2, -1, 5, and 6. Blood was collected and biopsied on d 6 of each period. Although we did not detect differences in milk yield, milk fat yield and content were lower in cows infused with DHA relative to PA. Plasma triacylglycerol concentrations were lower with DHA treatment relative to PA or BA. Cows infused with DHA had lower plasma insulin concentrations relative to cows infused with PA only. For objective 1, hepatic ceramide-d18:2/22:0 was highest in cows infused with BA relative to other treatments. For objective 2, plasma free choline concentrations were greater in PA+C cows relative to PA; however, we did not observe this effect with PA+S. Plasma total PC concentrations were similar for all treatments. Regarding the hepatic lipidome, a total of 18 hepatic PC were higher (e.g., PC-16:1/18:2) and 25 PC were lower (e.g., PC-16:0/22:6) with PA+C infusion relative to PA. In addition, 17 PC were higher (e.g., PC-20:3/22:5) and 21 PC were lower (e.g., PC-18:0/22:6) with PA+S infusion relative to PA. For objective 3, hepatic concentrations of many individual saturated PC (e.g., PC-18:0/15:0) were lower with DHA relative to other treatments. Hepatic concentrations of highly unsaturated PC with very-long-chain acids (e.g., PC-14:0/22:6) were higher in DHA-infused cows relative to PA, PA+C, PA+S, or BA. The abomasal infusion of emulsions containing , with choline chloride or serine, behenic , or docosahexaenoic influence the hepatic ceramide and PC profiles of lactating cows.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fatty liver

Celastrol reverses (PA)-caused TLR4-MD2 activation-dependent insulin resistance via disrupting MD2-related cellular binding to PA.

Elevated plasma statured acids (FFAs) cause TLR4/MD2 activation-dependent inflammation and insulin tolerance, which account for the occurrence and development of obesity. It has been confirmed that statured (PA) (the most abundant FFA) could bind MD2 to cause cellular inflammation. The natural compound celastrol could improve obesity, which is suggested via inhibiting inflammation, yet the detailed mechanism for celastrol is still unclear. As celastrol is reported to directly target MD2, we thought disrupting the binding between FFAs and MD2 might be one of the ways for celastrol to inhibit FFAs-caused inflammation and insulin resistance. In this study, we found evidence to support our hypothesis: celastrol could reverse PA-caused TLR4/MD2 activation-dependent insulin resistance, as determined by glucose-lowering ability, cellular glucose uptake, insulin action-related proteins and TLR4/MD2/NF-κB activation. Bioinformatics and cellular experiments showed that both celastrol and PA could bind MD2, and that celastrol could expel PA from cells. Finally, celastrol could reverse high fat diet caused hyperglycemia and obesity, and NF-kB activations. Taking together, we proved that celastrol could reverses PA-caused TLR4-MD2 activation-dependent insulin resistance via disrupting PA binding to MD2.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

Invited review: Sphingolipid biology in the dairy cow: The emerging role of ceramide.

The physiological control of lactation through coordinated adaptations is of fundamental importance for mammalian neonatal life. The putative actions of reduced insulin sensitivity and responsiveness and enhanced adipose tissue lipolysis spare glucose for the mammary synthesis of milk. However, severe insulin antagonism and body fat mobilization may jeopardize hepatic health and lactation in dairy cattle. Interestingly, lipolysis- and dietary-derived acids may impair insulin sensitivity in cows. The mechanisms are undefined yet have major implications for the development of postpartum disease. In nonruminants, the sphingolipid ceramide is a potent mediator of saturated fat-induced insulin resistance that defines in part the mechanisms of type 2 diabetes mellitus and nonalcoholic disease. In ruminants including the lactating dairy cow, the functions of ceramide had remained virtually undescribed. Through a series of hypothesis-centered studies, ceramide has emerged as a potential antagonist of insulin-stimulated glucose utilization by adipose and skeletal muscle tissues in dairy cattle. Importantly, bovine data suggest that the ability of ceramide to inhibit insulin action likely depends on the lipolysis-dependent hepatic synthesis and secretion of ceramide during early lactation. Although these mechanisms appear to fade as lactation advances beyond peak milk production, early evidence suggests that feeding is a means to augment ceramide supply. Herein, we review a body of work that focuses on sphingolipid biology and the role of ceramide in the dairy cow within the framework of hepatic and metabolism, insulin function, and lactation. The potential involvement of ceramide within the endocrine control of lactation is also considered.Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Keyword: fatty liver

Effects of Fat and Acids on the Formation of Autolysosomes in the Livers from Yellow Catfish Pelteobagrus Fulvidraco.

The autophagy-lysosome pathway, which involves many crucial genes and proteins, plays crucial roles in the maintenance of intracellular homeostasis by the degradation of damaged components. At present, some of these genes and proteins have been identified but their specific functions are largely unknown. This study was performed to clone and characterize the full-length cDNA sequences of nine key autolysosome-related genes ( and ) from yellow catfish Pelteobagrus fulvidraco. The expression of these genes and the transcriptional responses to a high-fat diet and acids (FAs) ( (PA) and oleic (OA)) were investigated. The mRNAs of these genes could be detected in heart, , muscle, spleen, brain, mesenteric adipose tissue, intestine, kidney and ovary, but varied with the tissues. In the , the mRNA levels of the nine autolysosome-related genes were lower in fish fed a high-fat diet than those fed the control, indicating that a high-fat diet inhibited formation of autolysosomes. (a saturated FA) significantly inhibited the formation of autolysosomes at 12 h, 24 h and 48 h incubation. In contrast, oleic (an unsaturated FA) significantly induced the formation of autolysosomes at 12 h, but inhibited them at 24 h. At 48 h, the effects of OA incubation on autolysosomes were OA concentration-dependent in primary hepatocytes of P. fulvidraco. The results of flow cytometry and laser confocal observations confirmed these results. PA and OA incubation also increased intracellular non-esterified (NEFA) concentration at 12 h, 24 h and 48 h, and influenced mRNA levels of binding protein () and transport protein 4 () which facilitate FA transport in primary hepatocytes of P. fulvidraco. The present study demonstrated the molecular characterization of the nine autolysosome-related genes and their transcriptional responses to fat and FAs in fish, which provides the basis for further exploring their regulatory mechanism in vertebrates.

Keyword: fatty liver

Palmitate and Stearate are Increased in the Plasma in a 6-OHDA Model of Parkinson\'s Disease.

Parkinson\'s disease (PD) is the second most common neurodegenerative disorder, without any widely available curative therapy. Metabolomics is a powerful tool which can be used to identify unexpected pathway-related disease progression and pathophysiological mechanisms. In this study, metabolomics in brain, plasma and was investigated in an experimental PD model, to discover small molecules that are associated with dopaminergic cell loss.Sprague Dawley (SD) rats were injected unilaterally with 6-hydroxydopamine (6-OHDA) or saline for the vehicle control group into the medial forebrain bundle (MFB) to induce loss of dopaminergic neurons in the substantia nigra pars compacta. Plasma, midbrain and samples were collected for metabolic profiling. Multivariate and univariate analyses revealed metabolites that were altered in the PD group.In plasma, ( = 3.72 × 10, FC = 1.81) and stearic ( = 3.84 × 10, FC = 2.15), were found to be increased in the PD group. ( = 3.5 × 10) and stearic ( = 2.7 × 10) correlated with test scores indicative of motor dysfunction. Monopalmitin ( = 4.8 × 10, FC = -11.7), monostearin ( = 3.72 × 10, FC = -15.1) and myo-inositol ( = 3.81 × 10, FC = -3.32), were reduced in the midbrain. The did not have altered levels of these molecules.Our results show that saturated free acids, their monoglycerides and myo-inositol metabolism in the midbrain and enteric circulation are associated with 6-OHDA-induced PD pathology.

Keyword: fatty liver

High vulnerability of the heart and to 3-hydroxypalmitic -induced disruption of mitochondrial functions in intact cell systems.

Patients affected by long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency predominantly present severe and cardiac dysfunction, as well as neurological symptoms during metabolic crises, whose pathogenesis is still poorly known. In this study, we demonstrate for the first time that pathological concentrations of 3-hydroxypalmitic (3HPA), the long-chain hydroxyl (LCHFA) that most accumulates in LCHAD deficiency, significantly decreased adenosine triphosphate-linked and uncoupled mitochondrial respiration in intact cell systems consisting of heart fibers, cardiomyocytes, and hepatocytes, but less intense in diced forebrain. 3HPA also significantly reduced mitochondrial Ca retention capacity and membrane potential in Ca -loaded mitochondria more markedly in the heart and the , with mild or no effects in the brain, supporting a higher susceptibility of the heart and the to the toxic effects of this . It is postulated that disruption of mitochondrial energy and Ca homeostasis caused by the accumulation of LCHFA may contribute toward the severe cardiac and hepatic clinical manifestations observed in the affected patients.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

A combination of plasma phospholipid acids and its association with incidence of type 2 diabetes: The EPIC-InterAct case-cohort study.

Combinations of multiple acids may influence cardiometabolic risk more than single acids. The association of a combination of acids with incident type 2 diabetes (T2D) has not been evaluated.We measured plasma phospholipid acids by gas chromatography in 27,296 adults, including 12,132 incident cases of T2D, over the follow-up period between baseline (1991-1998) and 31 December 2007 in 8 European countries in EPIC-InterAct, a nested case-cohort study. The first principal component derived by principal component analysis of 27 individual acids (mole percentage) was the main exposure (subsequently called the pattern score [FA-pattern score]). The FA-pattern score was partly characterised by high concentrations of linoleic , stearic , odd-chain acids, and very-long-chain saturated acids and low concentrations of γ-linolenic , , and long-chain monounsaturated acids, and it explained 16.1% of the overall variability of the 27 acids. Based on country-specific Prentice-weighted Cox regression and random-effects meta-analysis, the FA-pattern score was associated with lower incident T2D. Comparing the top to the bottom fifth of the score, the hazard ratio of incident T2D was 0.23 (95% CI 0.19-0.29) adjusted for potential confounders and 0.37 (95% CI 0.27-0.50) further adjusted for metabolic risk factors. The association changed little after adjustment for individual acids or subclasses. In cross-sectional analyses relating the FA-pattern score to metabolic, genetic, and dietary factors, the FA-pattern score was inversely associated with adiposity, triglycerides, enzymes, C-reactive protein, a genetic score representing insulin resistance, and dietary intakes of soft drinks and alcohol and was positively associated with high-density-lipoprotein cholesterol and intakes of polyunsaturated fat, dietary fibre, and coffee (p < 0.05 each). Limitations include potential measurement error in the acids and other model covariates and possible residual confounding.A combination of individual acids, characterised by high concentrations of linoleic , odd-chain acids, and very long-chain acids, was associated with lower incidence of T2D. The specific pattern may be influenced by metabolic, genetic, and dietary factors.

Keyword: fatty liver

Dual PPARα/γ agonist saroglitazar improves histopathology and biochemistry in experimental NASH models.

Non-alcoholic disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are common clinico-pathological conditions that affect millions of patients worldwide. In this study, the efficacy of saroglitazar, a novel PPARα/γ agonist, was assessed in models of NAFLD/NASH.HepG2 cells treated with (PA;0.75 mM) showed decreased expression of various antioxidant biomarkers (SOD1, SOD2, glutathione peroxidase and catalase) and increased expression of inflammatory markers (TNFα, IL1β and IL6). These effects were blocked by saroglitazar, pioglitazone and fenofibrate (all tested at 10μM concentration). Furthermore, these agents reversed PA-mediated changes in mitochondrial dysfunction, ATP production, NFkB phosphorylation and stellate cell activation in HepG2 and HepG2-LX2 Coculture studies. In mice with choline-deficient high-fat diet-induced NASH, saroglitazar reduced hepatic steatosis, inflammation, ballooning and prevented development of fibrosis. It also reduced serum alanine aminotransferase, aspartate aminotransferase and expression of inflammatory and fibrosis biomarkers. In this model, the reduction in the overall NAFLD activity score by saroglitazar (3\xa0mg/kg) was significantly more prominent than pioglitazone (25\xa0mg/kg) and fenofibrate (100\xa0mg/kg). Pioglitazone and fenofibrate did not show any improvement in steatosis, but partially improved inflammation and function. Antifibrotic effect of saroglitazar (4\xa0mg/kg) was also observed in carbon tetrachloride-induced fibrosis model.Saroglitazar, a dual PPARα/γ agonist with predominant PPARα activity, shows an overall improvement in NASH. The effects of saroglitazar appear better than pure PPARα agonist, fenofibrate and PPARγ agonist pioglitazone.© 2017 Cadila Healthcare Ltd., International Published by John Wiley & Sons Ltd.

Keyword: fatty liver

Strontium Alleviates Endoplasmic Reticulum Stress in a Nonalcoholic Disease Model.

The purpose of this study was to explore the effects of strontium on , and to clarify the possible mechanisms by which strontium improves nonalcoholic disease (NAFLD). We also evaluated how strontium affected the endoplasmic reticulum stress (ERS) pathways. We established an in vitro model of NAFLD using a human hepatocyte cell line (L02) treated with 0.2\u2009mM . The Sprague-Dawley rats were fed with a high-fat diet (HFD) to establish NAFLD model in vivo. After strontium treatment, the total cholesterol (TC), triglyceride (TG), and lipid deposition in L02 cells and tissues were determined. Strontium treatment suppressed intracellular TC and TG levels and lipid accumulation in L02 cells, and the effect of high concentrations of strontium were more obvious. Strontium significantly reduced the mRNA and protein expression of glucose-regulated protein 78 (GRP78), activating transcription factor 6 (ATF6), inositol requiring enzyme 1 (IRE1), SREBP cleavage activator protein (SCAP), sterol regulatory element binding protein 1c (SREBP-1c), and SREBP-2 in L02 cells. In HFD-fed rats, strontium treatment reduced serum TC, TG, and low density lipoprotein cholesterol (LDL-C) levels, concurrent with a decrease in hepatic lipid accumulation. Furthermore, strontium treatment reduced the expression of GRP78 and SREBP-2 protein in tissues. Overall, strontium alleviated hepatic steatosis by decreasing ERS-related protein expression in vivo and in vitro models. The results indicated that strontium has the potential to become a new therapy for the prevention and treatment of NAFLD.

Keyword: fatty liver

Individual acids in erythrocyte membranes are associated with several features of the metabolic syndrome in obese children.

Obesity leads to the clustering of cardiovascular (CV) risk factors and the metabolic syndrome (MetS) also in children and is often accompanied by non-alcoholic disease. Quality of dietary fat, beyond the quantity, can influence CV risk profile and, in particular, omega-3 acids (FA) have been proposed as beneficial in this setting. The aim of the study was to evaluate the associations of individual CV risk factors, characterizing the MetS, with erythrocyte membrane FA, markers of average intake, in a group of 70 overweight/obese children.We conducted an observational study. Erythrocyte membrane FA were measured by gas chromatography. Spearman correlation coefficients (r) were calculated to evaluate associations between FA and features of the MetS.Mean content of Omega-3 FA was low (Omega-3 Index\u2009=\u20094.7\u2009±\u20090.8%). Not omega-3 FA but some omega-6 FA, especially arachidonic (AA), were inversely associated with several features of the MetS: AA resulted inversely correlated with waist circumference (r\u2009=\u2009-\u20090.352), triglycerides (r\u2009=\u2009-\u20090.379), fasting insulin (r\u2009=\u2009-\u20090.337) and 24-h SBP (r\u2009=\u2009-\u20090.313). Total amount of saturated FA (SFA) and specifically , correlated positively with waist circumference (r\u2009=\u20090.354), triglycerides (r\u2009=\u20090.400) and fasting insulin (r\u2009=\u20090.287). Index (FLI), a predictive score of steatosis based on GGT, triglycerides and anthropometric indexes, was positively correlated to (r\u2009=\u20090.515) and inversely to AA (r\u2009=\u2009-\u20090.472).Our data suggest that omega-6 FA, and especially AA, could be protective toward CV risk factors featuring the MetS and also to indexes of hepatic steatosis in obese children, whereas SFA seems to exert opposite effects.

Keyword: fatty liver

Glucose and lipid metabolism disorders in the chickens with dexamethasone-induced oxidative stress.

The purpose of this study was to investigate the effects of long-term treatment with dexamethasone (DEX) on the antioxidation and nutrition metabolism in broiler chickens. Broilers were placed on a high-nutrient diet for 41\xa0days, and half were given orally DEX-supplemented water at 20\xa0mg/L every other day from 19 to 41\xa0days of age. DEX treatment downregulated superoxide dismutase activity as well as the mRNA expression of CuZn-superoxide dismutase and glutathione peroxidase with a decrease in GSH/GSSG ratio and an increase in malondialdehyde level in the of broilers. DEX treatment aggravated oxidative damage in the and, therefore, increased the sensitivity of broilers to ascites syndrome with higher mortality and reduced growth performance. Serum metabolomics analysis showed that DEX treatment significantly increased the levels of glucose, intermediates in protein metabolism (valine, proline, serine, threonine and urea) and lipid metabolism-related products (, stearic and cholesterol) while decreasing the levels of β-hydroxy butyric , succinic and malic , demonstrating that DEX treatment inhibited the Krebs cycle and the oxidation of acids, and promoted the de novo synthesis of acids as well as protein decomposition in the of broilers. Additionally, detection of metabolism-related enzymes\xa0revealed\xa0that DEX treatment inhibited glycolysis and promoted glycogen decomposition. In summary, DEX treatment resulted in oxidative stress and glucose and lipid\xa0metabolism disorders in the broilers.© 2017 Blackwell Verlag GmbH.

Keyword: fatty liver

Arazyme Suppresses Hepatic Steatosis and Steatohepatitis in Diet-Induced Non-Alcoholic Disease-Like Mouse Model.

Arazyme, a metalloprotease from the spider , exerts hepatoprotective activity in CCL-induced acute hepatic injury. This study investigated the hepatoprotective effects in high-fat diet (HFD)-induced non-alcoholic disease-like C57BL/6J mice. The mice were randomly divided into four groups ( = 10/group): the normal diet group, the HFD group, the arazyme group (HFD with 0.025% arazyme), and the milk thistle (MT) group (HFD with 0.1% MT). Dietary supplementation of arazyme for 13 weeks significantly lowered plasma triglyceride (TG) and non-esterified levels. Suppression of HFD-induced hepatic steatosis in the arazyme group was caused by the reduced hepatic TG and total cholesterol (TC) contents. Arazyme supplementation decreased hepatic lipogenesis-related gene expression, sterol regulatory element-binding transcription protein 1 (, synthase (), acetyl-CoA carboxylase 1 (), stearoyl-CoA desaturase-1 (), , glycerol-3-phosphate acyltransferase (), diacylglycerol -acyltransferase 1 (), and . Arazyme directly reduced (PA)-induced TG accumulation in HepG2 cells. Arazyme suppressed macrophage infiltration and tumor necrosis factor α (), interleukin-1β (), and chemokine-ligand-2 () expression in the , and inhibited secretion of TNFα and expression of inflammatory mediators, , , , , , and , in PA-induced RAW264.7 cells. Arazyme effectively protected hepatic steatosis and steatohepatitis by inhibiting SREBP-1-mediated lipid accumulation and macrophage-mediated inflammation.

Keyword: fatty liver

[Effects of sera of rats fed with tablets on endoplasmic reticulum stress in a HepG2 cell model of nonalcoholic disease].

To investigate the effects of sera from rats fed with tablets (HGT) on endoplasmic reticulum (ER) stress in a steatotic hepatocyte model of free acids (FFAs)-induced nonalcoholic disease (NAFLD) and explore the possible mechanism.FFAs prepared by mixing oleic and at the ratio of 2:1. HepG2 cells were treated with the sera from rats fed with low-, moderate-or high-dose HGT (HGT sera) or sera of rats fed with fenofibrate (fenofibrate sera), followed by treatment with 1 mmol/L FFAs for 24 h to induce hepatic steatosis. Oil red O staining was used to observe the distribution of lipid droplets in the cells. The biochemical parameters including triglyceride (TG), lactated hydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using a commercial kit. The morphological changes of the ER in the cells were observed using transmission electron microscopy. The protein/mRNA expressions of ER stress-related signal molecules including GRP78, PERK, p-PERK, ATF6, ATF4, CASPASE-12, CHOP, XBP-1, PKC, and p-PKC-δ were detected using Western blotting and/or quantitative real-time PCR (qRT-PCR). The changes in the protein expressions of GRP78, p-PERK, CASPASE-12 and CHOP were also detected in cells with transient transfection of PKC-δ siRNA for PKC-δ knockdown.Compared with the control cells, the cells treated with FFAs showed significantly increased levels of TG, AST, and ALT ( < 0.05). Compared with FFAs-treated cells, the cells pretreated with HGT sera or fenofibrate sera all showed significantly decreased TG, AST and ALT levels ( < 0.05), reduced accumulation of the lipid droplets ( < 0.05), and lowered protein or mRNA expression levels of GRP78, p-PERK, ATF6, ATF4, CHOP, CASPASE-12, XBP-1 and p-PKC-δ ( < 0.05). PKC-δ knockdown caused significantly reduced protein expressions of GRP78, p-PERK, CASPASE-12 and CHOP in the cells with FFA-induced hepatic steatosis ( < 0.001); treatment with high-dose HGT serum more significantly reduced the expressions of GRP78 ( < 0.001) and P-PERK ( < 0.01) in FFAs-induced cells with PKC-δ knockdown.HGT serum can effectively prevent FFAs-induced steatosis in HepG2 cells by alleviating ER stress, in which PKC-δ may act as an important target.

Keyword: fatty liver

Human Milk and Donkey Milk, Compared to Cow Milk, Reduce Inflammatory Mediators and Modulate Glucose and Lipid Metabolism, Acting on Mitochondrial Function and Oleylethanolamide Levels in Rat Skeletal Muscle.

Milk from various species differs in nutrient composition. In particular, human milk (HM) and donkey milk (DM) are characterized by a relative high level of triacylglycerol enriched in in sn-2 position. These dietary fats seem to exert beneficial nutritional properties through N-acylethanolamine tissue modulation. The aim of this study is to compare the effects of cow milk (CM), DM, and HM on inflammation and glucose and lipid metabolism, focusing on mitochondrial function, efficiency, and dynamics in skeletal muscle, which is the major determinant of resting metabolic rate. Moreover, we also evaluated the levels of endocannabinoids and N-acylethanolamines in and skeletal muscle, since tissue profiles can be modulated by nutrient intervention. To this aim, rats were fed with CM, DM, or HM for 4 weeks. Then, glucose tolerance and insulin resistance were analyzed. Pro-inflammatory and anti-inflammatory cytokines were evaluated in serum and skeletal muscle. Skeletal muscle was also processed to estimate mitochondrial function, efficiency, and dynamics, oxidative stress, and antioxidant/detoxifying enzyme activities. profiles, endocannabinoids, and N-acylethanolamine congeners were determined in and skeletal muscle tissue. We demonstrated that DM or HM administration reducing inflammation status, improves glucose disposal and insulin resistance and reduces lipid accumulation in skeletal muscle. Moreover, HM or DM administration increases redox status, and mitochondrial uncoupling, affecting mitochondrial dynamics in the skeletal muscle. Interestingly, HM and DM supplementation increase and muscle levels of the N-oleoylethanolamine (OEA), a key regulator of lipid metabolism and inflammation. HM and DM have a healthy nutritional effect, acting on inflammatory factors and glucose and lipid metabolism. This beneficial effect is associated to a modulation of mitochondrial function, efficiency, and dynamics and to an increase of OEA levels in skeletal muscle.

Keyword: fatty liver

Involvement of G protein-coupled receptor kinase 2 (GRK2) in the development of non-alcoholic steatosis and steatohepatitis in mice and humans.

Insulin resistance (IR) and obesity are important risk factors for non-alcoholic disease (NAFLD). G protein-coupled receptor kinase 2 (GRK2) is involved in the development of IR and obesity in vivo. However, its possible contribution to NAFLD and/or non-alcoholic steatohepatitis (NASH) independently of its role on IR or fat mass accretion has not been explored. Here, we used wild-type (WT) or GRK2 hemizygous (GRK2±) mice fed a high-fat diet (HFD) or a methionine and choline-deficient diet (MCD) as a model of NASH independent of adiposity and IR. GRK2± mice were protected from HFD-induced NAFLD. Moreover, MCD feeding caused an increased in triglyceride content and -to-body weight ratio in WT mice, features that were attenuated in GRK2± mice. According to their NAFLD activity score, MCD-fed GRK2± mice were diagnosed with simple steatosis and not overt NASH. They also showed reduced expression of lipogenic and lipid-uptake markers and less signs of inflammation in the . GRK2± mice preserved hepatic protective mechanisms as enhanced autophagy and mitochondrial fusion and biogenesis, together with reduced endoplasmic reticulum stress. GRK2 protein was increased in MCD-fed WT but not in GRK2± mice, and enhanced GRK2 expression potentiated -triggered lipid accumulation in human hepatocytes directly relating GRK2 levels to steatosis. GRK2 protein and mRNA levels were increased in human biopsies from simple steatosis or NASH patients in two different human cohorts. Our results describe a functional relationship between GRK2 levels and hepatic lipid accumulation and implicate GRK2 in the establishment and/or development of NASH.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

MiR-181b regulates steatosis in nonalcoholic disease via targeting SIRT1.

Non-alcoholic diseases (NAFLD) is one of the leading cause of chronic diseases in the world. However, the pathogenesis of NAFLD is still unclear. Emerging studies have demonstrated that microRNAs (miRs) are profoundly involved in NAFLD and related metabolic diseases. Here, we investigated the mechanisms by which miR-181b influences NAFLD via direct targeting SIRT1. The expression of miR181b was up-regulated while SIRT1 was down-regulated in both human NAFLD patients and high fat diet (HFD) induced NAFDL mice model. And (PA) treatment increased the miR-181b expression while decreased SIRT1 expression in HepG2 cells. Further, we identified that SIRT1 is a direct downstream target of miR-181b. Ectopic expression of miR-181b significantly repressed the 3\'-UTR reporter activities of SIRT1 in a dose-dependent manner, while the effect of miR-181b was interrupted when the binding site of miR-181b within the SIRT1 3\'-UTR was mutated. And overexpression of miR-181b reduced both the mRNA and protein levels of SIRT1 in HepG2 cells. We also found that inhibition of miR-181b expression alleviates hepatic steatosis both in\xa0vitro and in\xa0vivo. And the effect of miR-181b on steatosis was blocked by SIRT1 overexpression. Taken together, our data indicated that increased expression of miR-181b potentially contributes to altered lipid metabolism in NAFLD. Downregulation of miR-34a may be a therapeutic strategy against NAFLD by regulating its target SIRT1.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

[Insulin resistance is an alimentary deficiency of energy substrates (glucose) in the biological reaction of exotrophy and aphysiology compensation by acids via the biological reaction of endothrophy.]

The deficiency of energy substrates in the biological function of trophology and biological reaction of exotrophy is formed by two factors. Excess of meat in the diet leads to high content of (FA) in hepatocytes and formation of triglycerides (TG). Post heparin lipoprotein lipase slowly hydrolyzes TG in blood plasma lipoproteins and releases small amounts of FA. If dietary carbohydrate content is low, the biological function of exotrophy does not provide the substrate from which hepatocytes can rapidly produce oleic nonesterified FA de novo. Energy substrate deficiency activates the biological function of adaptation and the biological reaction of compensation. Under the effect of epinephrin NEFA deficiency is compensated via the biological reaction of endotrophy and lipolysis in omental visceral fat cells. In insulin resistance (IR) syndrome, the biological function of feeding is realized nonphysiologically while the biological reaction of adaptation is realized physiologically. An increase in NEFA blood content physiologically blocks glucose uptake in cells. Biological role of insulin consists in conversion of distant ocean-living carnivorous (fish-eating) ancestors of Homo sapiens with type of FA metabolism into herbivorous dry land-living species with oleic type metabolism of FA. The IR syndrome can be normalized. To this end a) the patient\'s will to activate the cognitive biological function (intellect) and b) comprehension of the fact that phylogenetically dry land-living Homo sapiens has developed as a herbivorous but not carnivorous species. Concerning death rate, cardiovascular pathologies are dominating in populations of many countries, while feeding function disorders prevail in frequency. These disorders form the pathophysiological basis for all metabolic pandemias: 1) atherosclerosis and atheromatosis, 2) essential arterial hypertension, 3) metabolic syndrome, 4) obesity, 5) insulin resistance syndrome, 6) nonalcoholic disease, and 7) endogenous hyperuricemia. Persistent potential deficiency of energy for realization of all biological reactions and functions is the major metabolic disorders in diabetes mellitus. Insulin resistance is a pathology associated primarily with FA and secondarily with glucose.

Keyword: fatty liver

Ellagic Suppresses the Oxidative Stress Induced by Dietary-Oxidized Tallow.

Dietary tallow was thermally oxidized at 180°C in an open fryer. The oxidized tallow (OT) and unoxidized tallow were characterized for oxidation parameters and composition using GC-MS. Tallow samples were fed to rabbits along with 50, 100, and 150\u2009mg/kg/day of ellagic (EA) for three weeks. Results revealed that the peroxide value (PV) and thiobarbituric reactive substances (TBARS) significantly increased, while radical scavenging activity (RSA) of the tallow decreased significantly with oxidation. GC-MS analysis showed eight acids in the tallow samples, where (48.5-49.7\u2009g/100\u2009g), linoleic (18.7-23.7\u2009g/100\u2009g), stearic (13.5-15.6\u2009g/100\u2009g), and margaric (6.32-6.42\u2009g/100\u2009g) were the major acids. Animal studies showed that oxidized tallow (OT) alone or in combination with EA significantly altered the body weight of the rabbits. Serum biochemical parameters and renal function tests were affected by OT and ameliorated by EA. The toxic effects of OT on haematological indices were minimized by EA. The supplementation of OT alone had significant effects on the structure and functions. The coadministration of EA reduced the toxic properties of OT on the , by increasing the antioxidant (GSH) system. The rabbit heart was also affected by the OT, which was ameliorated by EA supplementation. These results suggested that the supplementation of EA was beneficial against the OT-induced oxidative stress and may be considered for foods containing oxidized lipids.

Keyword: fatty liver

The effect of enterolactone on lipid precursors of inflammation.

The aim of this study was to assess the effects of enterolactone (ENL) on lipid fractions acids composition affecting hepatocyte inflammation development.The experiments were conducted in HepG2 cells incubated with ENL and/or (16\u202fh). Intracellular contents of free acids (FFA), di- (DAG) and tri- (TAG) acylglycerol as well as their acids compositions were assessed by Gas-Liquid Chromatography. Moreover, the ω-6/ω-3 ratios in the above mentioned lipids fractions were estimated. The expression of proteins involved in eicosanoids and prostanoids production (COX-2, 15-LOX), inflammatory process (TNFα), as well as the proteins participating in the desaturation (SCD 1) and elongation (Elovl 3, Elovl 6) of acids were evaluated by Western Blot.Enterolactone modified acids composition in FFA, DAG and TAG fractions. In conjunction with lipid overload, it increased the content of ω-6 more than ω-3 PUFA. Moreover, it enhanced the expressions of Elovl 3, Elovl 6, COX-2 and TNFα, whereas it had no influence on SCD 1 and 15-LOX level.Our study revealed that the supplementation with ENL affected intracellular hepatic composition of saturated as well as unsaturated acids in each of the investigated lipid fractions. Based on the shift in the ω-6/ω-3 balance towards ω-6, as well as the increase in COX-2 and TNFα protein expressions, we may postulate a pro-inflammatory nature of the examined polyphenol. Moreover, our findings could prove to be useful in the future research in the topic of widespread diseases such as NASH.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Downregulation of sirtuin 3 by increases the oxidative stress, impairment of mitochondrial function, and apoptosis in cells.

Elevated levels of saturated acids show a strong cytotoxic effect in cells. Sirtuin 3 (SIRT3), a mitochondrially localized member of NAD -dependent deacetylase has been shown to protect hepatocytes against the oxidative stress. The role of SIRT3 on the cytotoxicity caused by acids in cells is not fully understood. The aim of this study was to evaluate the expression level of SIRT3, oxidative stress, and mitochondrial impairments in human hepatoma HepG2 cells exposed to (PA). Our results showed that PA treatment caused the deposition of lipid droplets and resulted in an increased expression of tumor necrosis factor-α in a dose-dependent manner. Excessive accumulation of PA induces the reactive oxygen species formation and apoptosis while dissipating the mitochondrial transmembrane potential. The level of SIRT3 expression in both nuclear and mitochondrial fractions in HepG2 cells was decreased with the increase in PA concentrations. However, in the cytosolic fraction, the SIRT3 was undetectable. In conclusion, our results showed that PA caused an increase in inflammation and oxidative stress in HepG2 cells. The exposure of PA also resulted in the decline in transmembrane potential and an increase in apoptosis. The underexpression of nuclear and mitochondrial SIRT3 by PA suggests that the PA target the process that regulates the stress-related gene expression and mitochondrial functions.© 2019 Wiley Periodicals, Inc.

Keyword: fatty liver

[The person in philogenesis is not (omnivores), but the herbivores with the carnivores past and the fuzzy future. Biological function of trophology (nutrition) in ontogenesis.]

According to the phylogenetic theory of general pathology, seven biological functions have been formed over billions of years. 1. biological function of trophology, nutrition; 2. homeostasis function; 3. biological function of endoecology; 4. function of adaptation; 5. function of the continuation of the species; 6. function of locomotion and 7. cognitive biological function, including intelligence. Millions of years in life consistently in the waters of several oceans, all the ancestors of man were carnivorous (Carnivores), fish-eating mammals. When the ocean retreated and the carnivorous (fish-eating) were on land, each individual privatized a "piece" of the ocean. Animals transformed it ito a pool of intercellular medium in vivo. The biological role of the late in the phylogeny of insulin is the formation of new biological functions in vivo. The action of insulin has transformed the carnivorous (fish-eating) ocean into herbivorous (Herbivores) species on land. There was it by synthesis in vivo from exogenous glucose of acids (FA). Regulatory action of insulin was the directed conversion of exogenous glucose into ω-6 C18: 1 cis-oleic FA. Insulin late in phylogeny expressed the synthesis of new, conjugated enzymes: it is palmitoyl-CoA-elongase and stearyl-CoAdesaturase. Two enzymes synthesized FAs along the way: synthesized in situ de novo, from exogenous glucose, C16: 0 → C18: 0 stearic → ω-6 C18: 1 cis-oleic without accumulation of stearic FA. Insulin is not converted into an oleic FA exogenous from carnivorous food. On land, the action of insulin transformed the species Homo sapiens, into a herbivore, but with carnivorous, fish-eating, past. The idea of a person as omnivorous (Omnivor) - nonsense; such forms of nature did not form. Violation of the function of nutrition, the biological reaction of exotrophy (external nutrition), is the etiological and pathogenetic basis of the seven metabolic pandemics, the diseases of civilization. 1. Atherosclerosis and atheromatosis; 2. metabolic arterial hypertension; 3. metabolic syndrome; 4. obesity; 5. syndrome of insulin resistance; 6. non-alcoholic disease and 7. endogenous hyperuricemia. The primary prevention of metabolic pandemics in the biological function of nutrition, in the biological reactions of exo-and endotrophy, will allow us to understand the theoretical bases and implementation of preventive actions that will determine the characteristics of nutrition in the future.

Keyword: fatty liver

A Pathophysiological Model of Non-Alcoholic Disease Using Precision-Cut Slices.

Non-alcoholic disease (NAFLD) is a common disorder closely related to metabolic syndrome. NAFLD can progress to an inflammatory state called non-alcoholic steatohepatitis (NASH), which may result in the development of fibrosis and hepatocellular carcinoma. To develop therapeutic strategies against NAFLD, a better understanding of the molecular mechanism is needed. Current in vitro NAFLD models fail to capture the essential interactions between cell types and often do not reflect the pathophysiological status of patients. To overcome limitations of commonly used in vitro and in vivo models, precision-cut slices (PCLSs) were used in this study. PCLSs, prepared from tissue obtained from male Wistar rats, were cultured in supraphysiological concentrations of glucose, fructose, insulin, and to mimic metabolic syndrome. Accumulation of lipid droplets was visible and measurable after 24 h in PCLSs incubated with glucose, fructose, and insulin, both in the presence and absence of . Upregulation of acetyl-CoA carboxylase 1 and 2, and of sterol responsive element binding protein 1c, suggests increased de novo lipogenesis in PCLSs cultured under these conditions. Additionally, carnitine palmitoyltransferase 1 expression was reduced, which indicates impaired transport and disrupted mitochondrial β-oxidation. Thus, steatosis was successfully induced in PCLSs with modified culture medium. This novel ex vivo NAFLD model could be used to investigate the multicellular and molecular mechanisms that drive NAFLD development and progression, and to study potential anti-steatotic drugs.

Keyword: fatty liver

Permethrin and ivermectin modulate lipid metabolism in steatosis-induced HepG2 hepatocyte.

Recent studies have reported the positive association between exposure to insecticides and increased risk of obesity and type 2 diabetes, which are closely associated with non-alcoholic disease (NAFLD). However, it is not known if insecticide exposure can contribute to NAFLD. Thus, the goal of the current study was to determine if insecticide exposures can exacerbate the physiological conditions of NAFLD by modulating hepatic lipid metabolism. The effects of 12 insecticides on triglycerides (TG) accumulation were tested using (PA)-induced HepG2 hepatoma steatosis model. Results showed that among tested insecticides, permethrin and ivermectin significant interacted with to potentiate (permethrin) or decrease (ivermectin) TG accumulation. Further study showed that permethrin significantly promoted synthesis, while suppressed lipid oxidation-related genes only under steatosis conditions. In comparison, ivermectin inhibited lipogenesis-related genes and promoted farnesoid X receptor, which upregulates oxidation. Results in this study suggested that hepatic lipid metabolism may be more susceptible to insecticide exposure in the presence of excessive acids, which can be associated with the development of NAFLD.Copyright © 2019 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Reporting temporal fluctuations of hepatic C16 and C18 acids during late gestation and early lactation in dromedary camel.

Based on current knowledge, C16 and C18 acids (FA) are considered the most functional FA in hepatic metabolism. Although these FAs have been satisfyingly investigated in cattle, other species such as camel have been neglected. For this reason, the current study was designed to scrutinize changing patterns of C16 and C18 FAs in 10 dromedary camels from the last 2\xa0months of gestation to the first\xa0months of lactation. Camels were grazed on natural pasture and supplemented with a balanced ration. biopsies were obtained through blind biopsy technique at about 60, 45, 30, and 15-day antepartum (AP), and at 3, 15, 30, 45, and 60 post-partum (PP). Data were analyzed by the ANOVA procedure of SPSS with repeated measurements. From 15-day AP, saturated FA content of the declined (P\u2009<\u20090.01) and 15-day PP reached its peak (P\u2009=\u20090.02). At 30-day PP it went down (P\u2009<\u20090.01), and re-elevated at 45-day PP (P\u2009<\u20090.01) but remained at a steady state for the duration of the study. Mono-unsaturated and polyunsaturated FA content of hepatic tissue were constant throughout AP, albeit observed to peak at 15-day AP compared with 45 (P\u2009=\u20090.04) and 30-day AP (P\u2009<\u20090.01) for mono-unsaturated FAs, and with 60-, 45-, and 30-day AP (P\u2009≤\u20090.01) for polyunsaturated FAs. The content of the reached a nadir at 30-day AP (P\u2009<\u20090.01), increased sharply (P\u2009<\u20090.01) at the next sampling time-point, and had a trend to escalate until 3-day PP. Palmitoleic levels were unchanged from 60- to 30-day AP, decreased at 15 AP and 3-day PP, increased at 15-day PP, then remained constant until the end of the study period (P\u2009≤\u20090.04). Stearic content started to grow at 15-day AP and reached its peak at 15-day PP (P\u2009<\u20090.01). At 30-day PP, stearic level in dropped abruptly (P\u2009<\u20090.01), then intensified at 45-day PP and did not change after; hepatic content of stearic was lower during AP compared with PP time-points. Other C18 FAs changed significantly during the study period. These results suggest that parturition could have a profound effect on FA composition and other metabolites in camel . Further research is required to establish the metabolic mechanism behind these changes.

Keyword: fatty liver

GLP-1 Elicits an Intrinsic Gut- Metabolic Signal to Ameliorate Diet-Induced VLDL Overproduction and Insulin Resistance.

Perturbations in hepatic lipid and very-low-density lipoprotein (VLDL) metabolism are involved in the pathogenesis of obesity and hepatic insulin resistance. The objective of this study is to delineate the mechanism of subdiaphragmatic vagotomy in preventing obesity, hyperlipidemia, and insulin resistance.By subjecting the complete subdiaphragmatic vagotomized mice to various nutritional conditions and investigating hepatic de novo lipogenesis pathway, we found that complete disruption of subdiaphragmatic vagal signaling resulted in a significant decrease of circulating VLDL-triglyceride compared with the mice obtained sham procedure. Vagotomy further prevented overproduction of VLDL-triglyceride induced by an acute fat load and a high-fat diet-induced obesity, hyperlipidemia, hepatic steatosis, and glucose intolerance. Mechanistic studies revealed that plasma glucagon-like peptide-1 was significantly raised in the vagotomized mice, which was associated with significant reductions in mRNA and protein expression of SREBP-1c (sterol regulatory element-binding protein 1c), SCD-1 (stearoyl-CoA desaturase-1), and FASN ( synthase), as well as enhanced hepatic insulin sensitivity. In vitro, treating mouse primary hepatocytes with a glucagon-like peptide-1 receptor agonist, exendin-4, for 48 hours inhibited free , treatment induced de novo lipid synthesis, and VLDL secretion from hepatocytes.Elevation of glucagon-like peptide-1 in vagotomized mice may prevent VLDL overproduction and insulin resistance induced by high-fat diet. These novel findings, for the first time, delineate an intrinsic gut- regulatory circuit that is mediated by glucagon-like peptide-1 in regulating hepatic energy metabolism.© 2017 American Heart Association, Inc.

Keyword: fatty liver

Novel hypolipidemic conjugates of and bile with lysine for linkage.

Novel -bile conjugates (1a-1k) were designed and synthesized by coupling of the acids to the 3-OH of bile acids using lysine for linkage. In the conjugates, the 24-COOH of the bile acids was kept intact to preserve -specific recognition. The ability of the newly synthesized conjugates (at 100\u2009mg/kg dosage) to reduce total cholesterol (TC) and triglyceride (TG) levels in mice fed with high-fat diet (HFD) was evaluated. Conjugates of stearic with cholic and with ursodeoxycholic (at dosages of 50, 100, and 200\u2009mg/kg) were further evaluated to determine their ability to reduce aspartate aminotransferase (AST), alanine aminotransferase (ALT), TC, and TG levels in mice fed with HFD. All conjugates showed potent hypolipidemic activity. Further investigation revealed that compounds 1c and 1\u2009g not only dose-dependently reduced serum levels of TC and TG, but also inhibited the elevation of serum AST and ALT levels in mice fed with HFD. Thus, compounds 1c and 1\u2009g are promising hypolipidemic agents with hepatocyte protective effects against HFD-induced damage.

Keyword: fatty liver

Human plasma retinol-binding protein (RBP4) is also a -binding protein.

RBP4 (plasma retinol-binding protein) is the 21\u202fkDa transporter of all-trans retinol that circulates in plasma as a moderately tight 1:1 molar complex of the vitamin with the protein. RBP4 is primarily synthesized in the but is also produced by adipose tissue and circulates bound to a larger protein, transthyretin, TTR, that serves to increase its molecular mass and thus avoid its elimination by glomerular filtration. This paper reports the high resolution three-dimensional structures of human RBP4 naturally lacking bound retinol purified from plasma, urine and amniotic fluid. In all these crystals we found a molecule bound in the hydrophobic ligand-binding site, a result confirmed by mass spectrometry measurements. In addition we also report the 1.5\u202fÅ resolution structures of human holo-RBP4 and of the protein saturated with and lauric and discuss the interaction of the acids and retinol with the protein.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Vildagliptin Can Alleviate Endoplasmic Reticulum Stress in the Induced by a High Fat Diet.

We investigated whether a DDP-4 inhibitor, vildagliptin, alleviated ER stress induced by a high fat diet and improved hepatic lipid deposition. C57BL/6 mice received standard chow diet (CD), high fat diet (HFD), and HFD administered with vildagliptin (50\u2009mg/Kg) (V-HFD). After administration for 12 weeks, serum alanine aminotransferase, glucose, cholesterol, triglyceride, and insulin levels were analyzed. Samples of underwent histological examination and transmission electron microscopy, real-time PCR for gene expression levels, and western blots for protein expression levels. ER stress was induced in HepG2 cells with and the effects of vildagliptin were investigated. HFD mice showed increased weight/body weight (20.27%) and triglycerides (314.75%) compared to CD mice, but these decreased by 9.27% and 21.83%, respectively, in V-HFD mice. In the , HFD induced the expression of ER stress indicators significantly, which were obviously decreased by vildagliptin. In vitro, the expressions of molecular indicators of ER stress were reduced in HepG2 when vildagliptin was administered. Vildagliptin alleviates hepatic ER stress in a mouse high fat diet model. In HepG2 cells, vildagliptin directly reduced ER stress. Therefore, vildagliptin may be a potential agent for nonalcoholic disease.

Keyword: fatty liver

Long-term administration of tacrolimus and everolimus prevents high cholesterol-high fructose-induced steatosis in C57BL/6J mice by inhibiting de-novo lipogenesis.

To investigate the effects of tacrolimus (TC) and everolimus (EV) on non-alcoholic steatohepatitis (NASH) induced by high fat, high cholesterol and fructose (fast food) diet in C57BL/6J mice.C57BL/6J mice were divided into four groups (n=8). 1) Standard Chow (SC); 2) Fast food (FF) diet; 3) FF + Tacrolimus (TC, 1mg/kg) and; 4) FF + Everolimus (EV, 1mg/kg) and treated for 16 weeks. Serum and tissue samples were analyzed for evidence of inflammation, fibrosis, lipogenesis, and apoptosis.TC and EV treatments significantly reduced the hepatic lipid accumulation, improved -body weight ratio, blood biochemistry, and insulin resistance in mice fed with FF diet. However, inflammation, enlarged portal tracts, and fibrosis were pronounced in EV treated group. The lipogenic parameters, Peroxisome proliferator-activated receptor gamma (PPAR-γ), Sterol regulatory element-binding protein 1(SREBP-1), mammalian target of rapamycin (m-TOR), Stearoyl-CoA desaturase-1 (SCD-1) and translocase (CD36) were significantly down-regulated in livers of TC and EV treated groups as compared to FF group. TC improved Bcl2/Bax ratio, decreased apoptosis, CYP2E1 protein expression and fibrosis levels, however, EV offered no such protection. Further, in an model of lipotoxicity using the mouse hepatocyte (AML-12) cell line, treatment with TC and EV significantly reduced lipid accumulation and lipogenic and apoptotic markers induced with .In FF diet induced model of NASH, both TC and EV inhibited hepatic lipid accumulation and improved metabolic parameters such as insulin resistance and dyslipidemia. However, mice administered with EV exhibited inflammatory and fibrotic responses despite reduced hepatic steatosis.

Keyword: fatty liver

Dual Specificity Phosphatase 12 Regulates Hepatic Lipid Metabolism Through Inhibition of the Lipogenesis and Apoptosis Signal-Regulating Kinase 1 Pathways.

Nonalcoholic disease (NAFLD) has become the most common cause of chronic disease worldwide. Due to the growing economic burden of NAFLD on public health, it has become an emergent target for clinical intervention. DUSP12 is a member of the dual specificity phosphatase (DUSP) family, which plays important roles in brown adipocyte differentiation, microbial infection, and cardiac hypertrophy. However, the role of DUSP12 in NAFLD has yet to be clarified. Here, we reveal that DUSP12 protects against hepatic steatosis and inflammation in L02 cells after /oleic treatment. We demonstrate that hepatocyte specific DUSP12-deficient mice exhibit high-fat diet (HFD)-induced and high-fat high-cholesterol diet-induced hyperinsulinemia and steatosis and decreased insulin sensitivity. Consistently, DUSP12 overexpression in hepatocyte could reduce HFD-induced hepatic steatosis, insulin resistance, and inflammation. At the molecular level, steatosis in the absence of DUSP12 was characterized by elevated apoptosis signal-regulating kinase 1 (ASK1), which mediates the mitogen-activated protein kinase (MAPK) pathway and hepatic metabolism. DUSP12 physically binds to ASK1, promotes its dephosphorylation, and inhibits its action on ASK1-related proteins, JUN N-terminal kinase, and p38 MAPK in order to inhibit lipogenesis under high-fat conditions. Conclusion: DUSP12 acts as a positive regulator in hepatic steatosis and offers potential therapeutic opportunities for NAFLD.© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Diseases.

Keyword: fatty liver

Plasma lipidomic signatures of spontaneous obese rhesus monkeys.

Obesity plays crucial roles in the pathogenesis of metabolic diseases such as hyperlipidemia, nonalcoholic disease (NAFLD), and type 2 diabetes (T2D). The underlying mechanisms linking obesity to metabolic diseases are still less understandable.Previously, we screened a group of spontaneously obese rhesus monkeys. Here, we performed a plasma lipidomic analysis of normal and obese monkeys using gas chromatography/mass spectroscopy (GC/MS) and ultra-high performance liquid chromatography/mass spectroscopy (UPLC/MS).In total, 143 lipid species were identified, quantified, and classified into free acids (FFA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), lysophosphatidylcholine (LPC), lysophosphatidic (LPA), and sphingomyelin (SM). Data analysis showed that the obese monkeys had increased levels of acids palmitoleic (C16:1) and arachidonic (C20:4), FFA especially (C16:0), as well as certain PC species and SM species. Surprisingly, the plasma level of LPA-C16:0 was approximately four-fold greater in the obese monkeys. Conversely, the levels of most PE species were obviously reduced in the obese monkeys.Collectively, our work suggests that lipids such as FFA C16:0 and 16:0-LPA may be potential candidates for the diagnosis and study of obesity-related diseases.

Keyword: fatty liver

Ganoderma Lucidum Polysaccharide Peptide Alleviates Hepatoteatosis via Modulating Bile Metabolism Dependent on FXR-SHP/FGF.

Non-alcoholic disease (NAFLD) encompasses a series of pathologic changes ranging from steatosis to steatohepatitis, which may progress to cirrhosis and hepatocellular carcinoma. The purpose of this study was to determine whether ganoderma lucidum polysaccharide peptide (GLPP) has therapeutic effect on NAFLD.Ob/ ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD. Key metabolic pathways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blot. Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD.GLPP administrated for a month alleviated hepatosteatosis, dyslipidemia, dysfunction and insulin resistance. Pathways of glycerophospholipid metabolism, metabolism and primary bile biosynthesis were involved in the therapeutic effect of GLPP on NAFLD. Detection of key enzymes revealed that GLPP reversed low expression of CYP7A1, CYP8B1, FXR, SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice. Besides, GLPP inhibited synthesis by reducing the expression of SREBP1c, FAS and ACC via a FXR-SHP dependent mechanism. Additionally, GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepatocytes induced by oleic and .GLPP significantly improves NAFLD via regulating bile synthesis dependent on FXR-SHP/FGF pathway, which finally inhibits synthesis, indicating that GLPP might be developed as a therapeutic drug for NAFLD.© 2018 The Author(s). Published by S. Karger AG, Basel.

Keyword: fatty liver

Impact of Brain Signaling on Peripheral Insulin Action in Mice.

Glucose homeostasis and energy balance are under control by peripheral and brain processes. Especially insulin signaling in the brain seems to impact whole body glucose homeostasis and interacts with signaling. In humans circulating saturated acids are negatively associated with brain insulin action while animal studies suggest both positive and negative interactions of acids and insulin brain action. This apparent discrepancy might reflect a difference between acute and chronic signaling. To address this question we investigated the acute effect of an intracerebroventricular administration on peripheral glucose homeostasis. We developed and implemented a method for simultaneous monitoring of brain activity and peripheral insulin action in freely moving mice by combining radiotelemetry electrocorticography (ECoG) and euglycemic-hyperinsulinemic clamps. This method allowed gaining insight in the early kinetics of brain signaling and its contemporaneous effect on function , which, to our knowledge, has not been assessed so far in mice.Insulin-induced brain activity in the theta and beta band was decreased by acute intracerebroventricular application of . Peripherally it amplified insulin action as demonstrated by a significant inhibition of endogenous glucose production and increased glucose infusion rate. Moreover, our results further revealed that the brain effect of peripheral insulin is modulated by load in the brain.These findings suggest that insulin action is amplified in the periphery and attenuated in the brain by acute application. Thus, our results indicate that acute signaling in the brain may be different from chronic effects.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: fatty liver

Dietary lipid requirement of lemon fin barb hybrid.

The present study has been conducted to determine the optimal dietary requirement lipid of lemon fin barb hybrid fingerlings. Five isonitrogenous diets (35% protein) with graded lipid levels of 0%, 4%, 6%, 8% and 12% were fed to 225 lemon fin barb hybrid fingerlings (4.3 ? 0.5 g), which were randomly and equally distributed into twelve 60 l aquaria. The diets were randomly assigned to the aquaria in triplicates, and the feeding experiment was conducted for 10 weeks. Fish were fed at 5% body weight and the feeding ration was adjusted fortnightly. The weight gain, specific growth rate (SGR), protein efficiency ratio (PER) and feed conversion ratio (FCR) of fish fed 4% dietary lipid were significantly superior (P<0.05) than those fed with other diets, except for 6% lipid. Except for whole body moisture content, there were significant differences (P<0.05) between whole body proximate composition and nutrient retentions of fish fed with different test diets. The results showed that was the main saturated (SFA), oleic was the predominant MUFA while arachidic and docosahexanoic were the major PUFAs in the muscle and tissue of fish. The n-3/n-6 ratio ranged from 0.66 to 2.65. From the results of this study, a dietary lipid of 4% was recommended for lemon fin barb hybrid. ?

Keyword: fatty liver

Protective effect and mechanism of Qiwei Tiexie capsule on 3T3-L1 adipocytes cells and rats with nonalcoholic disease by regulating LXRα, PPARγ, and NF-κB-iNOS-NO signaling pathways.

Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic disease and nonalcoholic disease (NAFLD).This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD.The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and fat oxidation were assessed by 2-deoxy-D-[H] glucose and [1-C] , respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of fat emulsion and sucrose for 9 weeks. The effects of QWTX on lipid metabolism, function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the , as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected.QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the CO production rate to 4.15, which indicated the reducing the accumulation. In NAFLD, QWTX attenuated steatosis, fat vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model.QWTX protected the injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways.Copyright © 2019. Published by Elsevier B.V.

Keyword: fatty liver

[Psoralen and isopsoralen improve lipid metabolism disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic models were established by using ( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Keyword: fatty liver

In Vitro Monitoring of the Mitochondrial Beta-Oxidation Flux of and Investigation of Its Pharmacological Alteration by Therapeutics.

The present study was designed to validate the functional assay that enables rapid screening of therapeutic candidates for their effect on mitochondrial oxidation.The two whole-cell systems (tissue homogenates and hepatocytes) have been evaluated to monitor the total beta-oxidation flux of physiologically important H- by measurement of tritiated water enrichment in incubations using UPLC coupled on-line to radioactivity monitoring and mass spectrometry.Our results with several known inhibitors of oxidation showed that this simple assay could correctly predict a potential in alteration of mitochondrial function by drug candidates. Since the beta-oxidation of takes place almost exclusively in mitochondria of human hepatocytes, this model can be also utilized to distinguish between the mitochondrial and peroxisomal routes of this essential metabolic pathway in some cases.The present work offers a new in vitro screen of changes in mitochondrial beta-oxidation by xenobiotics as well as a model to study the mechanism of this pathway.

Keyword: fatty liver

Cypate and Cypate-Glucosamine as Near-Infrared Fluorescent Probes for In Vivo Tumor Imaging.

Near-infrared (NIR) imaging is a promising technique for use as a noninvasive and sensitive diagnostic tool. Although the NIR fluorescently labeled glucose analog glucosamine (cypate-glucosamine) has applications in preclinical imaging, the transport pathways and fate of this probe in tissues remain unaddressed. Here, we have synthesized and characterized cypate and cypate-glucosamine conjugate (cy-2-glu), and investigated the probable transport pathways of these probes in vitro and in vivo. We compared uptake of the probes in the presence and absence of excess d-glucose, "saturated cypate" and in two normal-cancer cell line pairs: lung cancer (A549)-normal (MRC9) and prostate cancer (DU145)-normal (BPH). Breast cancer (MDA-MB-231) and cancer (HepG2) cell lines were also examined. Results support use of the glucose transport pathway by cy-2-glu and transport pathway by cypate. Mass spectrometry data on the in vitro extracts revealed deamidation of cy-2-glu in prostate and cells, suggesting release of glucosamine. In vivo biodistribution studies in mice engrafted with breast tumors showed a distinct accumulation of cy-2-glu in and tumors, and to a lesser extent in kidneys and spleen. A negligible accumulation of cypate alone in tumors was observed. Analysis of urine extracts revealed renal excretion of the cy-2-glu probe in the form of free cypate, indicating deamidation of cy-2-glu in tissues. Thus, investigation of the metabolic pathways used by NIR probes such as cy-2-glu advances their use in the detection and monitoring of tumor progression in preclinical animal studies.Copyright © 2019 by The American Society for Pharmacology and Experimental Therapeutics.

Keyword: fatty liver

Thioacetamide potentiates high cholesterol and high fat diet induced steato-hepatitic changes in livers of C57BL/6J mice: A novel eight weeks model of fibrosing NASH.

There is an inadequacy of relevant animal models to study non-alcoholic steatohepatitis (NASH) and fibrosis. Here, we co-administered thioacetamide (TH) along with fast food diet (FFD) to C57BL/6\u2009J mice for eight weeks. The treatments were: a) standard chow, SC b) FFD c) FFD\u2009+\u2009TH [75\u2009mg/kg], FTH d) SC\u2009+\u2009TH [150\u2009mg/kg], STH for 8 weeks. In in-vitro model, Hep3B cells were exposed to (PA) and TH viz. PA (0.25\u2009mM) + TH (25\u2009mM), PA (0.5\u2009mM) alone and TH (50\u2009mM) alone for 12\u2009h, later supernatant media was transferred to LX-2 cells, for another 12\u2009h. Molecular and cellular events related to inflammation, fibrosis, collagen deposition were studied. The FTH mice featured hepatic inflammation, severe diffuse fibrosis, and collagen deposition, which were less severe in FF & STH groups. In FTH group the protein expressions of α-SMA, TGF-ß, Col1\u2009A1, CYP2E1, were up-regulated as compared to the FF group. The in-vivo findings were complemented in the LX-2 and Hep3B cells. The protein expressions of inflammatory and cellular injury markers were significantly higher in PA\u2009+\u2009TH exposed LX-2 cells. This novel model manifested hepatic inflammation and fibrosis in just eight weeks, which may be exploited for rapid screening of novel anti-NAFLD and anti-fibrotic agents.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Prepartum supplementation in sheep. IV. Effect of calcium salts with eicosapentaenoic and docosahexaenoic in the maternal and finishing diet on lamb and adipose tissue during the lamb finishing period1.

The objective of this study was to evaluate the effects of eicosapentaenoic (EPA) and docosahexaenoic (DHA) supplementation to ewes during late gestation on finishing lamb and adipose tissue (FA) profile and gene expression. Lambs born from ewes supplemented with Ca salts of EPA + DHA, or palm FA distillate (PFAD) high in and oleic at 0.39% DM during the last 50 d of gestation were used. Lambs were weaned at 61 d of age and adapted to a high concentrate diet for 1.5 mo. After adaptation, 74 lambs (28 pens) were blocked by sex and BW and used in a 2 × 2 factorial arrangement of treatments using the factors of dam supplementation (DS) and lamb supplementation (LS) of Ca salts of EPA + DHA or PFAD at 1.48% DM. Lambs were slaughtered after 42 d and and adipose tissue collected for FA and gene expression analysis. concentrations of EPA and DHA were greater (P < 0.01) with LS of EPA + DHA vs. PFAD during the finishing period. In adipose tissue, a lamb × dam interaction was observed for EPA (P = 0.02) and DHA (P = 0.04); LS of EPA + DHA increased EPA and DHA, but the increase was greatest in lambs born from ewes supplemented with PFAD. No lamb × dam treatment interactions were observed for gene expression in tissue (P > 0.10). Hepatic mRNA abundance of hormone-sensitive lipase (HSL; P = 0.01) was greater in lambs born from EPA + DHA ewes vs. lambs from PFAD ewes. mRNA expression of stearoyl-CoA desaturase (P < 0.01), synthase (P = 0.01), Δ5-desaturase (P < 0.01), and Δ6-desaturase (P < 0.01) were decreased in of EPA + DHA lambs. A significant lamb × dam diet interaction was observed for elongation of very long chain 2 in adipose tissue (P = 0.01); lambs supplemented with the same FA as their dams had lower expression. Expression of HSL tended (P = 0.08) to be decreased in adipose of EPA + DHA lambs born from EPA + DHA ewes. The changes in mRNA expression suggest that lipogenesis decreased, and lipolysis increased in lamb with EPA + DHA vs. PFAD supplementation during the finishing period. In adipose tissue, changes suggest that lipogenesis decreased in lambs born from EPA + DHA supplemented dams and supplemented with EPA + DHA during the finishing period. In addition, these results suggest an interaction between supplementation of FA to dams during late gestation on lamb response of adipose tissue, but not , to FA supplementation during the finishing period.© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Keyword: fatty liver

Aberrant hepatic lipid storage and metabolism in canine portosystemic shunts.

Non-alcoholic disease (NAFLD) is a poorly understood multifactorial pandemic disorder. One of the hallmarks of NAFLD, hepatic steatosis, is a common feature in canine congenital portosystemic shunts. The aim of this study was to gain detailed insight into the pathogenesis of steatosis in this large animal model. Hepatic lipid accumulation, gene-expression analysis and HPLC-MS of neutral lipids and phospholipids in extrahepatic (EHPSS) and intrahepatic portosystemic shunts (IHPSS) was compared to healthy control dogs. organoids of diseased dogs and healthy control dogs were incubated with - and oleic-, and lipid accumulation was quantified using LD540. In histological slides of shunt livers, a 12-fold increase of lipid content was detected compared to the control dogs (EHPSS P<0.01; IHPSS P = 0.042). Involvement of lipid-related genes to steatosis in portosystemic shunting was corroborated using gene-expression profiling. Lipid analysis demonstrated different triglyceride composition and a shift towards short chain and omega-3 acids in shunt versus healthy dogs, with no difference in lipid species composition between shunt types. All organoids showed a similar increase in triacylglycerols after free acids enrichment. This study demonstrates that steatosis is probably secondary to canine portosystemic shunts. Unravelling the pathogenesis of this hepatic steatosis might contribute to a better understanding of steatosis in NAFLD.

Keyword: fatty liver

Synthesis and evaluation of Ga labeled for cardiac metabolic imaging.

This work evaluates the potential of a Ga labeled long chain 16C for cardiac metabolic imaging. For radiolabeling with Ga, hexadecanedioic was coupled with the chelator p-NH-Bn-NOTA. Under the optimized conditions, NOTA-hexadecanoic could be radiolabeled with Ga in ≥95% yields. In biodistribution studies carried out in Swiss mice, Ga-NOTA-hexadecanoic showed low myocardial uptake at 2\u202fmin p.i. (3.7\u202f±\u202f1.3%ID/g). While Ga-NOTA-hexadecanoic cleared rapidly from non-target organs such as blood, lungs, intestine and kidney, wash out from was slow. Radio-HPLC analyses of myocardial extracts of rats injected with Ga-NOTA-hexadecanoic confirmed its metabolic transformation in the myocardium.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Berberine attenuates nonalcoholic hepatic steatosis through the AMPK-SREBP-1c-SCD1 pathway.

Berberine (BBR), a natural compound extracted from Chinese herb, has been shown to effectively attenuate nonalcoholic disease (NAFLD) in clinic. However, the mechanism underlying the effect of BBR is not fully understood. Stearyl-coenzyme A desaturase 1 (SCD1) mediates lipid metabolism in . Therefore, we hypothesized that SCD1 mediated the beneficial effect of BBR on NAFLD.The expression of SCD1 was measured in the of NAFLD patients and ob/ob mice. The effect of BBR on NAFLD was evaluated in C57BL/6\u202fJ mice on high fat diet (HFD). The effect of BBR was also investigated in HepG2 and AML12\u202fcells exposed to high glucose and . Oil red O staining was performed to detect triglyceride (TG) level. Quantitative real-time polymerase chain reaction and Western blot were used to detect the messenger ribonucleic (mRNA) and protein expression of target genes. The activity of SCD1 promoter was measured by dual-luciferase reporter assay.The expression of SCD1 was increased in the of NAFLD patients and ob/ob mice. BBR reduced hepatic TG accumulation and decreased the expressions of hepatic SCD1 and other TG synthesis related genes both in vivo and in vitro. Knockdown of SCD1 expression mimicked the effect of BBR decreasing TG level in steatotic hepatocytes, whereas overexpression of SCD1 attenuated the effect of BBR. Mechanistically, BBR promoted the phosphorylation of AMP-activated protein kinase (AMPK) and sterol regulatory element-binding protein-1c (SREBP-1c) in HepG2 cells and the of HFD-fed mice. Activation of the AMPK-SREBP-1c pathway and sterol regulatory element (SRE) motif in SCD1 promoter (-920/-550) was responsible for the BBR-induced suppression of SCD1.BBR reduces TG synthesis and attenuates hepatic steatosis through the activation of AMPK-SREBP-1c-SCD1 pathway.Copyright © 2019 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Hepatoprotective and Antioxidant Activities of Oil from Baru Almonds ( Vog.) in a Preclinical Model of Lipotoxicity and Dyslipidemia.

The oil obtained from baru ( Vog.) almonds exhibits high energy value and is reported in popular medicine for the treatment of rheumatic diseases and reproductive disturbances. Although baru oil is used in domestic cuisine, the chemical characterization of this oil and its effects on lipid metabolism are still poorly understood. Therefore, this study evaluated the (FA) profile and the effects of baru oil on and aorta in a murine model of dyslipidemia. The chromatographic profile of baru oil showed high levels of unsaturated FAs, especially oleic . Saturated FAs, such as and lignoceric acids, were found in lower amounts. Hypercholesterolemia was induced in male Wistar rats by daily administration of a lipid emulsion by gavage for 15 weeks. Biochemical and histopathological analysis were performed on serum, aorta, and . The results demonstrated that animals developed marked hypercholesterolemia, steatosis, and increased lipid peroxidation in the aorta. Treatment with baru oil attenuated lipid peroxidation and drastically reduced damage, especially ballooning degeneration and steatosis. By restricting vascular and hepatic injury, this oil showed potential applicability as a functional food, reinforcing its use in popular medicine and domestic cuisine.

Keyword: fatty liver

Inhibitory effect of 17β‑estradiol on triglyceride synthesis in skeletal muscle cells is dependent on ESR1 and not ESR2.

The present study aimed to investigate the inhibitory effects and the mechanisms underlying 17β‑estradiol (E2) effects on triglyceride synthesis and insulin resistance in skeletal muscle tissues and cells. Ovariectomy (OVX) was performed on 6‑month‑old female rats treated with or without E2. Subsequently, various serum biochemical markers were measured. Additionally, pathological alterations of the uterus, and skeletal muscle were analyzed, and the content of triglycerides (TG) in muscle was detected. Differentiated myotubes formed by C2C12 cells were treated with (PA) or pretreated with E2, estrogen receptor (ESR)\xa01 agonist propylpyrazoletriol (PPT) and ESR2 agonist diarylpropionitrile (DPN). Subsequently, the mRNA or protein expression levels of ESR1/2, peroxisome proliferator activated receptor α (PPARα), CD36 molecule (CD36), synthase (FASN), perilipin 2 (PLIN2), phosphorylated acetyl‑CoA carboxylase α (p‑ACACA), p‑AKT serine/threonine kinase (p‑AKT) and p‑mitogen‑activated protein kinase\xa08 (p‑MAPK8) were analyzed in skeletal muscle or in C2C12 cells by reverse transcription‑semi‑quantitative polymerase chain reaction and western blotting. The present results suggested that treatment with E2 inhibited OVX‑induced body weight gain, TG accumulation and insulin resistance. The protein or mRNA expression levels of ESR1, CD36, PPARα, p‑ACACA and p‑AKT were decreased, whereas the protein or mRNA expression levels of ESR2, PLIN2, FASN and p‑MAPK8 were increased in the OVX group. Of note, treatment with E2 restored the expression levels of the aforementioned factors. In C2C12 cells, treatment with E2 or PPT reversed the alterations induced by treatment with PA. In contrast, pretreatment with DPN did not influence the effect of PA. Collectively, E2 was able to interact with ESR1, thus activating the CD36‑PPARα pathway, decreasing the level of TG in the muscles and improving insulin resistance in skeletal muscles and C2C12 cells.

Keyword: fatty liver

NLRP3 Inflammasome Formation and Activation in Nonalcoholic Steatohepatitis: Therapeutic Target for Antimetabolic Syndrome Remedy FTZ.

The Nod-like receptor protein 3 (NLRP3) inflammasome activation not only serves as an intracellular machinery triggering inflammation but also produces uncanonical effects beyond inflammation such as changing cell metabolism and increasing cell membrane permeability. The present study was designed to test whether this NLRP3 inflammasome activation contributes to the "two-hit" injury during nonalcoholic steatohepatitis (NASH) and whether it can be a therapeutic target for the action of Fufang Zhenzhu Tiaozhi (FTZ), a widely used herbal remedy for hyperlipidemia and metabolic syndrome in China. We first demonstrated that NLRP3 inflammasome formation and activation as well as lipid deposition occurred in the of mice on the high-fat diet (HFD), as shown by increased NLRP3 aggregation, enhanced production of IL-1 and high mobility group box 1 (HMGB1), and remarkable lipid deposition in cells. FTZ extracts not only significantly reduced the NLRP3 inflammasome formation and activation but also attenuated the steatosis and fibrogenic phenotype changed. In studies, (PA) was found to increase colocalization of NLRP3 components and enhanced caspase-1 activity in hepatic stellate cells (HSCs), indicating enhanced formation and activation of NLRP3 inflammasomes by PA. PA also increased lipid deposition. Nlrp3 siRNA can reverse this effect by silencing the NLRP3 inflammasome and both with FTZ. In FTZ-treated cells, not only inflammasome formation and activation was substantially attenuated but also lipid deposition in HSCs was blocked. This inhibition of FTZ on lipid deposition was similar to the effects of glycyrrhizin, an HMGB1 inhibitor. Mechanistically, stimulated membrane raft redox signaling platform formation and increased O production by PA to activate NLRP3 inflammasomes in HSCs was blocked by FTZ treatment. It is concluded that FTZ extracts inhibit NASH by its action on both inflammatory response and lipid metabolism associated with NLRP3 inflammasome formation and activation.

Keyword: fatty liver

A unifying mathematical model of lipid droplet metabolism reveals key molecular players in the development of hepatic steatosis.

The responds to elevated plasma concentrations of free acids (FFAs) with an enhanced uptake of FFAs and their esterification to triacylglycerol (TAG). On the long term, this may result in massive hepatic TAG accumulation called steatosis hepatitis. In hepatocytes, the poor water-soluble TAG is packed in specialized organelles: Lipid droplets (LDs) serving as transient cellular deposit and lipoproteins (LPs) transporting TAG and cholesterol esters to extra-hepatic tissues. The dynamics of these organelles is controlled by a variety of regulatory surface proteins (RSPs). Assembly and export of VLDLs are mainly regulated by the microsomal transfer protein (MTP) and apoprotein B100. Formation and lipolysis of LDs are regulated by several RSPs. The best studied regulators belong to the PAT (Perilipin/Adipophilin/TIP47) and CIDE families. Knockdown or overexpression of SRPs may significantly affect the total number and size distribution of LDs. Intriguingly, a large cell-to-cell heterogeneity with respect to the number and size of LDs has been found in various cell types including hepatocytes. These findings suggest that the extent of cellular lipid accumulation is determined not only by the imbalance between lipid supply and utilization but also by variations in the expression of RSPs and metabolic enzymes. To better understand the relative regulatory impact of individual processes involved in the cellular TAG turnover, we developed a comprehensive kinetic model encompassing the pathways of the and triglyceride metabolism and the main molecular processes governing the dynamics of LDs. The model was parametrized such that a large number of experimental in vitro and in vivo findings are correctly recapitulated. A control analysis of the model revealed that variations in the activity of FFA uptake, diacylglycerol acyltransferase (DGAT) 2, and adipose triglyceride lipase (ATGL) have the strongest influence on the cellular TAG level. We used the model to simulate LD size distributions in human hepatoma cells and hepatocytes exposed to a challenge with FFAs. A random fold change by a factor of about two in the activity of RSPs was sufficient to reproduce the large diversity of droplet size distributions observed in individual cells. Under the premise that the same extent of variability of RSPs holds for the intact organ, our model predicts variations in the TAG content of individual hepatocytes by a factor of about 3-6 depending on the nutritional regime. Taken together, our modeling approach integrates numerous experimental findings on individual processes in the cellular TAG metabolism and LD dynamics metabolism to a consistent state-of-the-art dynamic network model that can be used to study how changes in the external conditions or systemic parameters will affect the TAG content of hepatocytes.© 2017 Federation of European Biochemical Societies.

Keyword: fatty liver

Increased plasma oleoylethanolamide and palmitoleoylethanolamide levels correlate with inflammatory changes in alcohol binge drinkers: the case of HMGB1 in women.

Alcohol binge drinking is a heavy pattern of alcohol consumption increasingly used by young people. In a previous study, we reported that young drinkers with a 2-year history of binge alcohol consumption had an overactivation of the innate immune system and peripheral inflammation when compared with controls. In the present study, we measured several biolipids that are derivatives belonging to the acylethanolamide or 2-acylglycerol families in the plasma of the same subjects (n\xa0=\xa042; 20 men and 22 women). We found that during abstinence, alcohol binge drinkers had elevated plasma levels of oleoylethanolamide, palmitoleoylethanolamide, arachidonoylethanolamide, dihomo-γ-linolenoyl ethanolamide and linoleoyl ethanolamide, which positively correlated with changes in the mRNA expression of key inflammatory markers in peripheral blood mononuclear cells, such as toll-like receptors (TLR4), pro-inflammatory cytokines/chemokines interleukin-1 beta, interleukin-6 and monocyte chemoattractant protein-1, and cyclooxygenase-2. Additionally, plasma oleoylethanolamide positively correlated with plasma levels of high mobility group box-1, which is a danger-associated molecular pattern and an endogenous TLR4 agonist, specifically in female alcohol binge drinkers. No changes were observed in 2-acylglycerols in alcohol binge drinkers, although sex-related differences in these bioactive lipids as well as in palmitoleoylethanolamide and docosatetraenoylethanolamide levels were detected. These results extend the previous clinical findings observed in patients diagnosed with long-term alcohol use disorder to young users and suggest a prominent role for these lipids in the response to acute alcohol exposure.© 2017 Society for the Study of Addiction.

Keyword: fatty liver

Palmitate induces nitric oxide production and inflammatory cytokine expression in zebrafish.

Inflammation markers in zebrafish embryos reflect a toxic response that is common to other animal models and humans. Free acids (FFAs) are known to cause damage in various tissues by inducing inflammation. In this study, we investigated whether a FFA (palmitate) induces inflammation in zebrafish embryos. Nitrous oxide (NO) production and cyclooxygenase-2 (COX-2) mRNA expression were increased in palmitate-treated zebrafish embryos in a dose-dependent manner. mRNA expression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF- α), were also increased. Additionally, the mRNA expression of p65 nuclear factor-kB and I-kB-α were significantly increased after palmitate-treatment. Increased reactive oxygen species (ROS) expression was observed in palmitate-treated zebrafish embryos as well as pericardial edema. Additionally, mRNA expression of pro-inflammatory cytokines were increased in zebrafish and pancreas fed with palmitate-contained diet. Taken together, these results indicated that palmitate increases pro-inflammatory mediators in zebrafish embryos, suggesting that zebrafish could be an alternative animal model for inflammatory disease including diabetes.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Hepatitis C Virus Infection Increases c-Jun N-Terminal Kinase (JNK) Phosphorylation and Accentuates Hepatocyte Lipoapoptosis.

BACKGROUND Hepatitis C virus (HCV) infection and metabolic diseases including nonalcoholic steatohepatitis (NASH) exhibit a complex interplay. Although free -mediated apoptosis is a prominent feature of NASH, the impact of HCV infection on hepatocyte lipotoxicity has remained largely unexplored. The study aimed at identifying whether infection by HCV affected the apoptotic pathway in hepatocytes during assault. MATERIAL AND METHODS OR6 cells, which are derived from human hepatocellular carcinoma Huh-7 cells and harbor a full-length HCV RNA genome replication system, were treated with palmitate. Apoptosis was examined by 4\',6-diamidino-2-phenylindole staining. Activation and expression of JNK, Bim, cIAP-1, and Mcl-1 were examined by immunoblotting. mRNA expression of CHOP, a major player in endoplasmic reticulum stress-mediated apoptosis, was assessed by real-time PCR. RESULTS Palmitate-induced hepatocyte apoptosis was significantly enhanced in OR6 cells compared to cured cells, in which the HCV genome had been eradicated by treatment with interferon-α. Although basal expression of CHOP mRNA was enhanced in OR6 cells compared to cured cells, it was similarly upregulated in both cell lines following palmitate treatment. Notably, palmitate-induced JNK phosphorylation was accentuated in OR6 cells compared to cured cells. Inhibition of JNK with SP600125 attenuated palmitate-induced apoptosis. Palmitate-mediated upregulation of BH3-only protein Bim, which acts downstream of JNK, was also enhanced in OR6 cells compared to cured cells. In contrast, Mcl-1 and cIAP-1 were equally reduced in OR6 cells and cured cells following palmitate treatment. CONCLUSIONS These findings suggest that during lipoapoptosis, HCV infection may enhance hepatocyte toxicity by increasing JNK phosphorylation.

Keyword: fatty liver

TRIF-dependent Toll-like receptor signaling suppresses transcription in hepatocytes and prevents diet-induced hepatic steatosis.

Nonalcoholic disease (NAFLD) includes a spectrum of diseases that ranges in severity from hepatic steatosis to steatohepatitis, the latter of which is a major predisposing factor for cirrhosis and cancer. Toll-like receptor (TLR) signaling, which is critical for innate immunity, is generally believed to aggravate disease progression by inducing inflammation. Unexpectedly, we found that deficiency in TIR domain-containing adaptor-inducing interferon-β (TRIF), a cytosolic adaptor that transduces some TLR signals, worsened hepatic steatosis induced by a high-fat diet (HFD) and that such exacerbation was independent of myeloid cells. The aggravated steatosis in mice was due to the increased hepatocyte transcription of the gene encoding stearoyl-coenzyme A (CoA) desaturase 1 (SCD1), the rate-limiting enzyme for lipogenesis. Activation of the TRIF pathway by polyinosinic:polycytidylic [poly(I:C)] suppressed the increase in SCD1 abundance induced by or an HFD and subsequently prevented lipid accumulation in hepatocytes. Interferon regulatory factor 3 (IRF3), a transcriptional regulator downstream of TRIF, acted as a transcriptional suppressor by directly binding to the promoter. These results suggest an unconventional metabolic function for TLR/TRIF signaling that should be taken into consideration when seeking to pharmacologically inhibit this pathway.Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Keyword: fatty liver

Pioglitazone Enhances Cytosolic Lipolysis, β-oxidation and Autophagy to Ameliorate Hepatic Steatosis.

Non-alcoholic disease closely contributes to the development of obesity and insulin resistance. Even though pioglitazone has been reported to effectively lessen hepatic steatosis in human studies, its molecular mechanism remains unclear. This study is designed to investigate the regulation of cytosolic lipolysis, β-oxidation and autophagy by pioglitazone in a mice model of high fat diet (HFD) and cell model incubated with . Our results revealed hepatic steatosis was apparently induced by HFD and it was significantly reversed by pioglitazone. The serum insulin and hepatic triglyceride content was significantly decreased by co-administered pioglitazone with HFD. Hepatic expression of cytosolic-lipolysis related proteins (ATGL, HSL), β-oxidation (CPT-1A) and autophagy-related proteins (ATG7, LC3, LAL) was significantly enhanced by pioglitazone. Knockdown PPARα/PPARγ in AML12 cells significantly and proportionally reduced the expressions of ATGL, CPT-1A and LC3II, which was induced by pioglitazone. Furthermore, facilitation of the autophagic flux by pioglitazone was obviously blocked by lysosomal inhibitor, leupeptin, to demonstrate accumulation of the LC3II and intracellular lipid in AML12 cells. Our results demonstrated that pioglitazone attenuating the hepatic steatosis may be mediated by enhancing cytosolic lipolysis, β-oxidation and autophagy in a PPARα and PPARγ dependent manner.

Keyword: fatty liver

Matrine attenuates endoplasmic reticulum stress and mitochondrion dysfunction in nonalcoholic disease by regulating SERCA pathway.

Endoplasmic reticulum (ER) stress, which can promote lipid metabolism disorders and steatohepatitis, contributes significantly to the pathogenesis of nonalcoholic disease (NAFLD). Calcium (Ca) homeostasis is considered to play a key role in ER stress. Matrine (Mat) has been applied for the treatment of hepatitis B, but its effect on NAFLD is still unknown, and there is no unified view of Mat on the regulation of ER stress in the previous literature.The pharmacological effects were studied in high-fat-diet or methionine-choline-deficient diet induced C57BL/6J mice models and in (PA) induced L02 human cell model. Calcium fluorescence experiments, computational virtual docking analysis and biochemical assays were used in identifying the locus of Mat.The results showed that Mat-treated mice were more resistant to steatosis in the than vehicle-treated mice and that Mat significantly reduced hepatic inflammation, lipid peroxides. The beneficial effect of Mat was associated with suppressing ER stress and restoring mitochondrial dysfunction. Additionally, Mat decreased the PA-induced lipid accumulation, ER stress and cytosolic calcium level ([Ca]) in hepatocyte cell lines in low and middle dose. However, the high dose Mat did not show satisfactory results in cell model. Calcium fluorescence experiments showed that Mat was able to regulate [Ca]. By computational virtual docking analysis and biochemical assays, Mat was shown to influence [Ca] via direct inhibition of SERCA.The results showed that the bi-directional regulation of Mat to endoplasmic reticulum at different doses was based on the inhibition of SERCA. In addition, the results also provide a theoretical basis for Mat as a potential therapeutic strategy in NAFLD/NASH.

Keyword: fatty liver

Polydatin attenuates diet-induced nonalcoholic steatohepatitis and fibrosis in mice.

Non-alcoholic steatohepatitis (NASH) is characterized by lipid accumulation in hepatocytes and inflammatory cell infiltration. In view of the anti-oxidative and anti-inflammatory effects of polydatin, the current study aimed to investigate the pharmacological effects of polydatin on NASH and its related fibrosis. C57BL/6 mice were fed with methionine-choline deficient (MCD) diet to induce NASH and fibrosis, and treated with or without polydatin (5 mg/kg, every other day, i.p) for 4 weeks. HepG2 cells induced by (PA) were treated with polydatin. The elevations of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), active caspase-3, TUNEL-positive cells, and triglyceride content were decreased by polydatin treatment. In addition, administration of polydatin to MCD-fed mice reduced oxidative stress by down-regulating NOX4 enzymes. Furthermore, the reduction in inflammation and CD68 macrophage activation correlated with inhibition of toll-like receptor (TLR)-4/NF-κB p65 signaling pathway by polydatin treatment. Polydatin also attenuated lipid accumulation, inflammation and apoptosis in HepG2 cells challenged by (PA) combined with or without lipopolysaccharide (LPS). Finally, the reduction of hepatic fibrosis by polydatin treatment corresponded to a reduction in hepatic gene expression of fibrosis markers. These results suggest that polydatin prevents NASH and fibrosis via inhibition of oxidative stress and inflammation, highlighting polydatin as a potential therapeutic agent for prevention and treatment of NASH.

Keyword: fatty liver

GC-MS metabolomics reveals disturbed metabolic pathways in primary mouse hepatocytes exposed to subtoxic levels of 3,4-methylenedioxymethamphetamine (MDMA).

3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is a well-known hepatotoxic drug. Although its toxicity has been thoroughly studied at high concentrations, there is still insufficient knowledge on possible alterations of cell function at subtoxic concentrations, which are in fact more representative concentrations of intoxication scenarios. In this study, a gas chromatography-mass spectrometry (GC-MS) metabolomics approach was used to investigate the metabolic changes in primary mouse hepatocytes (PMH) exposed to two subtoxic concentrations of MDMA (LC and LC) for 24\xa0h. Metabolomic profiling of both intracellular metabolites and volatile metabolites in the extracellular medium of PMH was performed. Multivariate analysis showed that the metabolic pattern of cells exposed to MDMA discriminates from the controls in a concentration-dependent manner. Exposure to LC MDMA induces a significant increase in some intracellular metabolites, including oleic and , and a decrease in glutamate, aspartate, 5-oxoproline, fumarate, malate, phosphoric , α-ketoglutarate and citrate. Extracellular metabolites such as acetophenone, formaldehyde, pivalic , glyoxal and 2-butanone were found significantly increased after exposure to MDMA, compared to controls, whereas 4-methylheptane, 2,4-dimethyl-1-heptene, nonanal, among others, were found significantly decreased. The panel of discriminatory metabolites is mainly involved in tricarboxylic (TCA) cycle, metabolism, glutamate metabolism, antioxidant defenses and possibly changes in the enzyme machinery. Overall, these results highlight the potential of the intra- and extracellular metabolome to study alterations triggered by subtoxic concentrations of MDMA in hepatic cell functions, which represents a more realistic appraisal of early toxicity events posed by exposure to this drug. In addition, these results also revealed some metabolites that may be used as potential biomarkers indicative of early events in the hepatotoxicity induced by MDMA.

Keyword: fatty liver

Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARα/PPARγ coactivator-1α pathway.

Nuciferine, an alkaloid found in Nelumbo nucifera leaves, alleviates dyslipidemia in vivo. However, whether it improves injury in diabetic conditions and the underlying mechanism is unclear. The present study aimed to investigate the effects of nuciferine on lipid and glucose metabolism in a murine model of Type 2 diabetes mellitus (T2DM) and to determine the underlying mechanisms of these effects.A murine model of T2DM was induced by high-fat diet (HFD) feeding combined with streptozocin (STZ) injections, and the diabetic mice were treated with nuciferine in their food. The underlying mechanism of the anti-steatotic effect of nuciferine was further explored in HepG2 hepatocytes cultured with . Major signalling profiles involved in oxidation were then evaluated, using Western blot, RT-qPCR and si-RNA techniques, along with immunohistochemistry.Nuciferine restored impaired glucose tolerance and insulin resistance in diabetic mice. Hepatic levels of total cholesterol, triglycerides and LDL were decreased, as were the number of lipid droplets, by nuciferine treatment. Furthermore, nuciferine up-regulated β-oxidation related genes in livers of diabetic mice. Luciferase reporter cell assay showed that nuciferine directly reversed -induced inhibition of PPARα transcriptional activity. Silencing PPARγ coactivator-1α (PGC1α) expression in HepG2 cells abolished the effects of nuciferine in accelerating β-oxidation.Nuciferine improved lipid profile and attenuated hepatic steatosis in HFD/STZ-induced diabetic mice by activating the PPARα/PGC1α pathway. Nuciferine may be a potentially important candidate in improving hepatic steatosis and the management of T2DM.© 2018 The British Pharmacological Society.

Keyword: fatty liver

Free -induced histone acetyltransferase activity accelerates lipid accumulation in HepG2 cells.

Non-alcoholic disease (NAFLD) is a common metabolic disease triggered by epigenetic alterations, including lysine acetylation at histone or non-histone proteins, affecting the stability or transcription of lipogenic genes. Although various natural dietary compounds have anti-lipogenic effects, their effects on the acetylation status and lipid metabolism in the have not been thoroughly investigated.Following oleic- (OPA)-induced lipid accumulation in HepG2 cells, the acetylation status of histone and non-histone proteins, HAT activity, and mRNA expression of representative lipogenic genes, including γ, , , and , were evaluated. Furthermore, correlations between lipid accumulation and HAT activity for 22 representative natural food extracts (NExs) were evaluated.Non-histone protein acetylation increased following OPA treatment and the acetylation of histones H3K9, H4K8, and H4K16 was accelerated, accompanied by an increase in HAT activity. OPA-induced increases in the mRNA expression of lipogenic genes were down-regulated by C-646, a p300/CBP-specific inhibitor. Finally, we detected a positive correlation between HAT activity and lipid accumulation (Pearson\'s correlation coefficient = 0.604) using 22 NExs.Our results suggest that NExs have novel applications as nutraceutical agents with HAT inhibitor activity for the prevention and treatment of NAFLD.

Keyword: fatty liver

Novel Fluorescence-Based Method To Characterize the Antioxidative Effects of Food Metabolites on Lipid Droplets in Cultured Hepatocytes.

A fluorescence microscopic method for characterizing size, quantity, and oxidation of lipid droplets (LDs) in HepG2 cells was developed. LDs were induced by (PA), oleic (OA), or linoleic acids (LA) and stained with two fluorescent probes for neutral lipids and lipid peroxides. Each fatty increased the number of LDs and oxidized LDs (oxLDs) and the degree of LD oxidation time dependently, as well as increased intracellular triglyceride hydroperoxides. LDs induced by LA without 2,2\'-azobis(2-amidinopropane)dihydrochloride (AAPH) showed the most significant oxidation degree over PA and OA, especially in large LDs (area ≥ 3 μm, oxLD/LD = 52.3 ± 21.7%). Under this condition, two food-derived antioxidants were evaluated, and both of them significantly improved the LD characteristics. Moreover, chlorogenic reduced the quantity of large LDs by 74.0-87.6% in a dose-dependent manner. The proposed method provides a new approach to evaluate the effect of dietary antioxidants on LD characteristics.

Keyword: fatty liver

Alleviation of -induced endoplasmic reticulum stress by augmenter of regeneration through IP3R-controlled Ca release.

The aberrant release of Ca from the endoplasmic reticulum (ER) contributes to the onset of ER stress, which is closely related to the pathogenesis of non-alcoholic disease. We previously reported that augmenter of regeneration (ALR) alleviates ER stress and protects hepatocytes from lipotoxicity. However, the link between ALR protection and the suppression of ER stress remains unclear. In this study, we investigated whether the protection against steatosis afforded by ALR is related to its inhibition of calcium overflow from the ER to the mitochondria. The treatment of HepG2 cells with (PA) upregulated IP3R expression, triggering ER-luminal Ca release and inducing ER stress. However, in ALR-transfected (ALR-Tx) HepG2 cells, PA-induced cell injury was clearly alleviated compared with that in vector-Tx cells. After exposure to PA, IP3R expression was downregulated and ER stress was effectively inhibited in the ALR-Tx cells, and ER-Ca release and simultaneous mitochondrial Ca uptake were lower than those in vector-Tx cells. The knockdown of ALR expression with shRNA abolished the protective effects afforded by ALR transfection. PA treatment also suppressed the interaction between BCL-2 and IP3R in HepG2 cells, whereas this interaction was massively enhanced in the ALR-Tx cells, effectively reducing the IP3R-mediated ER-Ca release and thus mitochondrial Ca influx. Our results suggest that the inhibition of ER stress by ALR is related to the interruption of the interaction between BCL2 and IP3R, demonstrating a novel mechanism of ER stress resistance in ALR-Tx cells.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

Low molecular weight fucoidan attenuates injury via SIRT1/AMPK/PGC1α axis in db/db mice.

Non-alcoholic disease (NAFLD), caused by elevated hepatic lipids, inflammation and oxidative stress, is the most common disease globally. Low molecular weight fucoidan (LMWF), a sulfated polysaccharide extracted from brown seaweeds, has shown strong anti-inflammatory and antioxidant activities, which has not been explored in diabetes-induced NAFLD. Therefore, the present study sought to determine whether LMWF protects obese diabetic db/db mice against NAFLD. Results showed LMWF administration decreased plasma level of alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride, as well as alleviated hepatic accumulation of triglyceride and total cholesterol in db/db mice. LMWF also ameliorated hepatic oxidative stress by suppressing superoxide production and lipid peroxidation, and increasing catalase and superoxide dismutase activity in the of db/db mice. Furthermore, LMWF down-regulated several pro-inflammatory cytokines and transcription factor, and up-regulated the anti-inflammatory adiponectin. These changes were accompanied by the activation of hepatic SIRT1/AMPK/PGC1α signaling with LMWF treatment. In addition, blocking SIRT1 or AMPK by inhibitor notably abolished LMWF-elicited protection against -induced oxidative stress and inflammation in hepatocytes. These results suggest LMWF prevents NAFLD in db/db mice by activation of SIRT1/AMPK/PGC1α signaling pathway, which prevents lipotoxicity-related oxidative stress and inflammation. Therefore, LMWF provides a potential supplementary treatment for obesity/diabetes-induced NAFLD.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Attenuation of -Induced Lipotoxicity by Chlorogenic through Activation of SIRT1 in Hepatocytes.

Saturated free acids (FFAs) induce hepatocyte lipotoxicity, wherein oxidative stress-associated mitochondrial dysfunction is mechanistically involved. Chlorogenic (CGA), a potent antioxidant and anti-inflammatory compound, protects against high-fat-diet-induced oxidative stress and mitochondrial dysfunction in . This study investigates whether CGA protects against FFA-induced hepatocyte lipotoxicity via the regulation of mitochondrial fission/fusion and elucidates its underlying mechanisms.AML12 cell, a non-transformed hepatocyte cell line, is treated with palmitate. Here, it is shown that CGA prevents palmitate-induced lipotoxicity by activation of SIRT1 regulated mitochondrial morphology. CGA treatment mitigates oxidative stress and mitochondrial dysfunction, as evidenced by a decrease in reactive oxygen species (ROS) production, and an increase in mitochondrial mass and mitochondrial membrane potential. CGA also significantly decreases Bax expression and thereby reduces mitochondria-mediated caspase-dependent apoptosis. Mechanistically, CGA attenuates ROS-induced mitochondrial fragmentation by inhibiting dynamin-related protein 1 (Drp1) and enhancing Mfn2 expression. In contrast, the inhibitory effects of CGA on the generation of mitochondrial ROS and Drp1 are blocked by siRNA knockdown of SIRT1.Collectively, these findings show that supplementation with CGA protects hepatocytes from FFA-induced lipotoxicity through activation of SIRT1, which reverses the oxidative stress and dysfunction of mitochondrial biogenesis directly.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fatty liver

The effect of enterolactone on sphingolipid pathway and hepatic insulin resistance development in HepG2 cells.

Obesity and type 2 diabetes mellitus, correlate with increased tissue concentration of sphingolipids, which directly interfere with insulin signaling pathway. Phytoestrogens are a group of plant-derived compounds that have been studied in the case of metabolic disorders treatment. Therefore, the aim of this study was to ascertain whether enterolactone (ENL), a commonly known phytoestrogen, may affect sphingolipid metabolism and decrease hepatic insulin resistance development in a lipid overload state.The study was conducted on HepG2 cells incubated with ENL and/or (PA) for 16\u202fh. Intra- and extracellular sphingolipid concentrations were assessed by high performance liquid chromatography. The expression of sphingolipid pathway enzymes, apoptosis and insulin signaling pathway proteins and glucose metabolism regulators were evaluated by Western Blot.In HepG2 cells, a considerable augmentation of intracellular ceramide and sphingosine concentration in ENL with PA group were indicated with simultaneous increase in extracellular ceramide concentration. The ENL treatment increased expression of selected enzymes from de novo ceramide synthesis pathway with lower expression of ceramide transfer protein. We also observed a decreased expression of insulin-stimulated phosphorylation of AKT and AMPK after exposure to ENL with PA. Our research demonstrated that ENL with PA resulted in an increased expression of caspase-3.Enterolactone, in a higher acids availability, led to the development of hepatic IR in HepG2 cells. This phenomenon may be the result of elevated intracellular ceramide accumulation caused by increased de novo synthesis pathway what led to enhanced apoptosis of HepG2 cells.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Oleic protects saturated mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis.

Aim This study aims to demonstrate the protective effects of monounsaturated oleic (OA) against saturated (PA) induced cellular lipotoxicity in hepatocytes and rats with non-alcoholic steatohepatitis (NASH).Human hepatoma cell line HepG2 cells and neonatal rat primary hepatocytes were treated with PA or/and OA for 24\u202fh. SD rats were fed with high fat diet (HFD) to induce NASH. From the 16th w, the HFD was full or half replaced by olive oil to observe the protective effects.In vitro, OA substantially alleviated PA induced cellular apoptosis, oxidative stress, ER stress, mitochondrial dysfunction, as well as inflammation in hepatocytes. In vivo, only olive oil supplementation had no detrimental effects, while HFD developed NASH in normal rats. Full replacement of HFD with olive oil had profoundly reversed NASH. Noteworthily, half replacement of HFD with olive oil (a mixed diet) has ameliorated NASH injury as well. It strikingly changed the hepatic histology from macrovesicular-steatosis into entire microvesicular-steatosis, and significantly reduced inflammation, ballooning and fibrosis.Our study has demonstrated in both hepatocytes and NASH rats that oleic acids had great potential to combat the saturated acids induced hepatic lipotoxicity. Only half replacement of HFD by monounsaturated acids rich diet still had significant therapeutic outcome in NASH rats. Redirecting the toxic saturated acids into triglyceride storage and reduction of cholesterol accumulation might be the possible explanation of OA driven protection in this scenario.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Silibinin ameliorates hepatic lipid accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway.

Non-alcoholic steatohepatitis (NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic plus , and adenovirus-down for 24\u202fh, then treated with silibinin for 24\u202fh. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of acids in to relieve lipid accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

Keyword: fatty liver

Acids Consumption: The Role Metabolic Aspects Involved in Obesity and Its Associated Disorders.

Obesity and its associated disorders, such as insulin resistance, dyslipidemia, metabolic inflammation, dysbiosis, and non-alcoholic hepatic steatosis, are involved in several molecular and inflammatory mechanisms that alter the metabolism. Food habit changes, such as the quality of acids in the diet, are proposed to treat and prevent these disorders. Some studies demonstrated that saturated acids (SFA) are considered detrimental for treating these disorders. A high fat diet rich in , a SFA, is associated with lower insulin sensitivity and it may also increase atherosclerosis parameters. On the other hand, a high intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids may promote positive effects, especially on triglyceride levels and increased high-density lipoprotein (HDL) levels. Moreover, polyunsaturated acids (PUFAs) and monounsaturated acids (MUFAs) are effective at limiting the hepatic steatosis process through a series of biochemical events, such as reducing the markers of non-alcoholic hepatic steatosis, increasing the gene expression of lipid metabolism, decreasing lipogenic activity, and releasing adiponectin. This current review shows that the consumption of unsaturated acids, MUFA, and PUFA, and especially EPA and DHA, which can be applied as food supplements, may promote effects on glucose and lipid metabolism, as well as on metabolic inflammation, gut microbiota, and hepatic metabolism.

Keyword: fatty liver

Serum lipids as an indicator for the alteration of function in patients with hepatitis B.

Hepatitis B virus (HBV) exerts an intense impact on host lipid metabolism. Hence the aim of present study is to determine metabolic derangement that occurred in subjects suffering from hepatitis B patients.The fasting blood samples were collected from hepatitis B patients (n\u2009=\u200950) attended in Taluka hospital TandoAdam, Sindh with age and gender matched controls (n\u2009=\u200950). Serum lipid profile and (FA) composition were analyzed by micro-lab and gas chromatography.The hepatitis B patients have significantly lower level (p\u2009<\u20090.01) of lipid profile including total cholesterol (TC), triacylglyceride (TAG), high density lipoprotein-C (HDL-C) very low density lipoprotein-cholesterol (VLDL-C), low density lipoprotein-cholesterol (LDL-C), and total lipid (TL) in comparison to controls, indicating hypolipidemia in patients. The result of total FA composition of HBV patients in comparison to controls reveal that myristic, , docosahexaenoic acids were significantly (p\u2009<\u20090.05) higher, while linoleic, eicosatrienoic, arachidonic, eicosapentaenoic acids were lower in HBV patients in comparison to controls. The elongase, ∆5 and ∆6-desaturase enzymes activities were found lower, while ∆9-desaturase activity was higher in hepatitis B patients as compared to controls, which indicates the impaired lipid metabolism.The serum saturated (SFA) and monounsaturated (MUFA) were increased while polyunsaturated (PUFA) was reduced in both total and free form in hepatitis B patients due to altered activities of enzyme desaturases with impaired PUFA metabolism and non-enzymatic oxidation.

Keyword: fatty liver

High α-tocopherol dosing increases lipid metabolism by changing redox state in damaged rat gastric mucosa and after ethanol treatment.

Regeneration of ethanol-injured rat gastric mucosa must undergo changes in major metabolic pathways to achieve DNA replication and cell proliferation. These events are highly dependent on glucose utilization and inhibited by vitamin E (VE) (α-tocopherol) administration. Therefore, the present study aimed at assessing lipid metabolism in the gastric mucosa and ethanol-induced gastric damage and the effect of α-tocopherol administration. For this, rates of β-oxidation and lipogenesis were tested in gastric mucosa samples. Through histological analysis, we found loss of the mucosa\'s superficial epithelium, which became gradually normalized during the recovery period. Proliferation of gastric mucosa occurred with augmented formation of β-oxidation by-products, diminished synthesis of triacylglycerols (TGs), as well as of phospholipids, and a reduced cytoplasmic NAD/NADH ratio, whereas the mitochondrial redox NAD/NADH ratio was much less affected. In addition, α-tocopherol increased utilization in the gastric mucosa, which was accompanied by the induction of \'mirror image\' effects on the cell redox state, reflected in an inhibited cell gastric mucosa proliferation by the vitamin administration. In conclusion, the present study shows, for the first time, the role of lipid metabolism in the adaptive cell gastric mucosa changes that drive proliferation after a chronic insult. Moreover, α-tocopherol increased gastric mucosa utilization of associated with energy production. These events could be associated with its antioxidant properties in co-ordination with regulation of genes and cell pathways, including changes in the cell NAD/NADH redox state.© 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Keyword: fatty liver

Saturated combined with lipopolysaccharide stimulates a strong inflammatory response in hepatocytes in vivo and in vitro.

Nonalcoholic disease (NAFLD) is the most common chronic disease and consumption of high-fat diet (HFD) is a risk factor for NAFLD. The HFD not only increases intake of saturated (SFA) but also induces metabolic endotoxemia, an HFD-associated increase in circulating lipopolysaccharide (LPS). Although it is known that SFA or LPS promote hepatic inflammation, a hallmark of NAFLD, it remains unclear how SFA in combination with LPS stimulates host inflammatory response in hepatocytes. In this study, we performed both in vivo and in vitro experiments to investigate the effect of SFA in combination with LPS on proinflammatory gene expression in hepatocytes. Our animal study showed that feeding low-density lipoprotein-deficient mice HFD enriched with SFA and injection of low-dose LPS cooperatively stimulated IL-6 expression in livers. To understand how SFA and LPS interact to promote IL-6 expression, our in vitro studies showed that (PA), a major SFA, and LPS exerted synergistic effect on the expression of IL-6 in hepatocytes. Furthermore, coculture of hepatocytes with macrophages resulted in a greater IL-6 expression than culture of hepatocytes without macrophages in response to the combination of PA and LPS. Finally, we observed that LPS and PA increased ceramide production by cooperatively stimulating ceramide de novo synthesis, which played an essential role in the synergistic stimulation of proinflammatory gene expression by LPS and PA. Taken together, this study showed that SFA in combination with LPS stimulated a strong inflammatory response in hepatocytes in vivo and in vitro.

Keyword: fatty liver

Increased expression of sterol regulatory element binding protein‑2 alleviates autophagic dysfunction in NAFLD.

Sterol regulatory element binding protein‑2 (SREBP‑2) is an important transcription factor in lipid homeostasis. A previous study showed that SREBP‑2 also activated autophagic genes during cell‑sterol depletion. Alterations in autophagy are reported to be involved in the pathogenesis of nonalcoholic disease (NAFLD). However, whether the regulation of SREBP‑2 restores dysfunctional autophagy in hepatocytes during NAFLD remains to be elucidated. In the present study, a steatosis model was established with (PA) treatment at the indicated times and concentrations. Autophagosomes in hepatocytes were visualized by confocal microscopy after transfection with a tandem GFP‑mCherry‑LC3 construct. Autophagy‑associated protein levels were analyzed by western blot analysis. Loss‑ and gain‑of‑function studies were performed to examine the role of SREBP‑2 in the regulation of hepatocyte autophagy. It was demonstrated that PA induced autophagy and enhanced autophagic flux at the early stage, whereas prolonged treatment with PA resulted in dysfunction of autophagy in the PA‑induced steatotic hepatocytes. In addition, different cellular models presented with differing dysfunctional autophagy in response to overload. It was also confirmed that SREBP‑2 regulated autophagy‑related gene expression in hepatocytes, and it was shown that the overexpression of SREBP‑2 increased the expression of autophagy‑related genes, but did not affect the inhibition of the autophagic flux in lipid‑overloaded HL‑7702 cells. By contrast, increased SREBP‑2 partly restored the inhibited autophagic activity in lipid‑overloaded hepatoma HepG2 cells. Taken together, the present study demonstrated that autophagic function was impaired in lipid‑overloaded human hepatocytes, and the differential effect of PA on autophagy was associated with the duration of PA and the cell type. Under these conditions, the overexpression of SREBP‑2 alleviated the inhibited autophagic activity rather than the inhibition of autophagic flux. Consequently, the results indicated that restoration of autophagy dysfunction via the regulation of SREBP‑2 may be a potential therapeutic target for the treatment of NAFLD.

Keyword: fatty liver

Impaired mitophagy triggers NLRP3 inflammasome activation during the progression from nonalcoholic to nonalcoholic steatohepatitis.

Activation of inflammation is an important mechanism in the development of nonalcoholic steatohepatitis (NASH). This study aims to delineate how mitophagy affects NLRP3 inflammasome activation in hepatic lipotoxicity. Mice were fed a high fat/calorie diet (HFCD) for 24 weeks. Primary rat hepatocytes were treated with (PA) for various periods of time. Mitophagy was measured by protein levels of LC3II and P62. NLRP3, caspase-1, interleukin (IL)-18, and IL-1β at mRNA and protein levels were used as indicators of inflammasome activation. Along with steatotic progression in HFCD-fed mice, ratio of LC3II/β-actin was decreased concurrently with increased levels of P62, NLRP3, caspase-1, IL-1β, IL-18, and serum IL-1β levels in late-stage NASH. PA treatment resulted in mitochondrial oxidative stress and initiated mitophagy in primary hepatocytes. The addition of cyclosporine A did not change LC3II/Τοmm20 ratios; but P62 levels were increased after an extended duration of PA exposure, indicating a defect in autophagic activity. Along with impaired mitophagy, mRNA and protein levels of NLRP3, caspase-1, IL-18 and IL-1β were upregulated by PA treatment. Pretreatment with MCC950, N-acetyl cysteine or acetyl-L-carnitine reversed inflammasome activation and a pyroptotic cascade. Additionally, mitophagic flux was partially recovered as indicated by increases in LC3II/Tomm20 ratio, parkin, and PINK1 expression, and decreased P62 expression. The findings suggest that impaired mitophagy triggers hepatic NLRP3 inflammasome activation in a murine NASH model and primary hepatocytes. The new insights into inflammasome activation through mitophagy advance our understanding of how acids elicit lipotoxicity through oxidant stress and autophagy in mitochondria.

Keyword: fatty liver

Genomics of lipid-laden human hepatocyte cultures enables drug target screening for the treatment of non-alcoholic disease.

Non-alcoholic disease (NAFLD) is a major health burden in need for new medication. To identify potential drug targets a genomic study was performed in lipid-laden primary human hepatocyte (PHH) and human hepatoma cell cultures.PHH, HuH7 and HepG2 hepatoma cell cultures were treated with lipids and/or TNFα. Intracellular lipid load was quantified with the ORO assay. The Affymetrix HG-U133+ array system was employed to perform transcriptome analysis. The lipid droplet (LD) growth and fusion was determined by fluorescence microscopy. LD associated proteins were imaged by confocal immunofluorescence microscopy and confirmed by Western immunoblotting. Bioinformatics defined perturbed metabolic pathways.Whole genome expression profiling identified 227, 1031 and 571 significant regulated genes. Likewise, the combined lipid and TNFα treatment of PHH, HuH7 and HepG2 cell cultures revealed 154, 1238 and 278 differentially expressed genes. Although genomic responses differed among in-vitro systems, commonalities were ascertained by filtering the data for LD associated gene regulations. Among others the LD-growth and fusion associated cell death inducing DFFA like effector C (CIDEC), perilipins (PLIN2, PLIN3), the synaptosome-associated-protein 23 and the vesicle associated membrane protein 3 were strongly up-regulated. Likewise, the PPAR targets pyruvate-dehydrogenase-kinase-4 and angiopoietin-like-4 were up-regulated as was hypoxia-inducible lipid droplet-associated (HILPDA), flotilin and FGF21. Their inhibition ameliorates triglyceride and cholesterol accumulation. TNFα treatment elicited strong induction of the chemokine CXCL8, the kinases MAP3K8, MAP4K4 and negative regulators of cytokine signaling, i.e. SOCS2&SOCS3. Live cell imaging of DsRED calreticulin plasmid transfected HuH7 cells permitted an assessment of LD growth and fusion and confocal immunofluorescence microscopy evidenced induced LD-associated PLIN2, CIDEC, HIF1α, HILPDA, JAK1, PDK4 and ROCK2 expression. Notwithstanding, CPT1A protein was repressed to protect mitochondria from lipid overload. Pharmacological inhibition of the GTPase-dynamin and the transporter-2 reduced lipid uptake by 28.5 and 35%, respectively. Finally, a comparisons of in-vitro/NAFLD patient biopsy findings confirmed common gene regulations thus demonstrating clinical relevance.The genomics of fat-laden hepatocytes revealed LD-associated gene regulations and perturbed metabolic pathways. Immunofluorescence microscopy confirmed expression of coded proteins to provide a rationale for therapeutic intervention strategies. Collectively, the in-vitro system permits testing of drug candidates.

Keyword: fatty liver

Hepatocyte miR-33a mediates mitochondrial dysfunction and hepatosteatosis by suppressing NDUFA5.

Emerging evidence suggests that microRNAs (miRNAs) are essential for metabolic haemostasis of tissues. Among them, miR-33a is supposed to modulate the cholesterol export and oxidation, but whether miR-33a involves in the process of disease is unclear. To disclose the hypothesis, we utilized miR-33a mimic and antisense to explore their effects in primary hepatocytes or high-fat diet (HFD)-fed mice. Treatment with (PA) or HFD significantly increased the expression of miR-33a in hepatocytes or tissues. In primary hepatocytes, miR-33a mimic decreased mitochondrial function, including reduction of ATP production and oxygen consumption, whereas miR-33a inhibition protected PA-induced mitochondrial dysfunction. Interestingly, miR-33a selectively suppressed mitochondrial complex I activity and protein expression, but not other complexes. Through bioinformatics prediction, we found miR-33a directly targeted on the 3\'-UTR of NDUFA5. Dual-luciferase reporter analysis further confirmed the direct suppression of miR-33a on NDUFA5 expression. More importantly, administration of miR-33a antisense could effectively restore HFD-induced mitochondrial dysfunction through up-regulation of NDUFA5 levels. Mice treated with miR-33a antisense also exhibited improved function and structural disorders under obese status. Taken together, miR-33a was an important mediator of hepatocyte mitochondrial function, and the therapeutic benefits implied miR-33a antisense had the potential clinical application in combating the disease.© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fatty liver

Dihydromyricetin Ameliorates Nonalcoholic Disease by Improving Mitochondrial Respiratory Capacity and Redox Homeostasis Through Modulation of SIRT3 Signaling.

Our previous clinical trial indicated that the flavonoid dihydromyricetin (DHM) could improve hepatic steatosis in patients with nonalcoholic disease (NAFLD), altough the potential mechanisms of these effects remained elusive. Here, we investigated the hepatoprotective role of DHM on high-fat diet (HFD)-induced NAFLD. DHM supplementation could effectively ameliorate the development of NAFLD by inhibiting hepatic lipid accumulation both in HFD-fed wild-type mice and in -induced hepatocytes. We reveal for the first time that mitochondrial dysfunction characterized by ATP depletion and augmented oxidative stress could be reversed by DHM treatment. Moreover, DHM enhanced the mitochondrial respiratory capacity by increasing the expression and enzymatic activities of mitochondrial complexes and increased mitochondrial reactive oxygen species scavenging by restoring manganese superoxide dismutase (SOD2) activity. Interestingly, the benefits of DHM were abrogated in knockout (SIRT3KO) mice and in hepatocytes transfected with siRNA or treated with an SIRT3-specific inhibitor. We further showed that DHM could increase SIRT3 expression by activating the adenosine monophosphate-activated protein kinase (AMPK)-peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC1α)/estrogen-related receptor-α (ERRα) signaling pathway. Our work indicates that SIRT3 plays a critical role in the DHM-mediated beneficial effects that include ameliorating mitochondrial dysfunction and oxidative stress in a nutritional NAFLD model both and Our results suggest that DHM prevents NAFLD by improving mitochondrial respiratory capacity and redox homeostasis in hepatocytes through a SIRT3-dependent mechanism. These results could provide a foundation to identify new DHM-based preventive and therapeutic strategies for NAFLD.

Keyword: fatty liver

N-3 Polyunsaturated Acids Decrease Long-Term Diabetic Risk of Offspring of Gestational Diabetes Rats by Postponing Shortening of Hepatic Telomeres and Modulating Metabolism.

The long-term influence of gestational diabetes mellitus (GDM) on offspring and the effect of omega-3 polyunsaturated acids (n-3 PUFA) on GDM offspring are poorly understood. We studied the long-term diabetic risk in GDM offspring and evaluated the effect of n-3 PUFA intervention. Healthy offspring rats were fed standard diet (soybean oil) after weaning. GDM offspring were divided into three groups: GDM offspring (soybean oil), n-3 PUFA adequate offspring (fish oil), and n-3 PUFA deficient offspring (safflower oil), fed up to 11 months old. The diabetic risk of GDM offspring gradually increased from no change at weaning to obvious impaired glucose and insulin tolerance at 11 months old. N-3 PUFA decreased oxidative stress and inflammation in the of older GDM offspring. There was a differential effect of n-3 PUFA and n-6 PUFA on hepatic telomere length in GDM offspring. Non-targeted metabolomics showed that n-3 PUFA played a modulating role in the , in which numerous metabolites and metabolic pathways were altered when GDM offspring grew to old age. Many metabolites were related to diabetes risk, such as α-linolenic , , ceramide, oxaloacetic , tocotrienol, tetrahydro-11-deoxycortisol, andniacinamide. In summary, GDM offspring exhibited obvious diabetes risk at old age, whereas n-3 PUFA decreased this risk.

Keyword: fatty liver

[The involvement of NOX1/NADPH oxidase in the development of non-alcoholic steatohepatitis].

Reactive oxygen species (ROS) are known to play a critical role in the development of non-alcoholic steatohepatitis (NASH). To clarify the source of ROS, we examined the expression of superoxide-generating NADPH oxidase isoforms in the of high-fat and high-cholesterol (HFC) diet-fed mice. The mRNA expression of NOX1 was significantly elevated in mice on HFC diet for 8 weeks. Increased levels of serum alanine aminotransferase and hepatic cleaved caspase-3 in HFC diet-fed wild-type mice (WT) were significantly ameliorated in mice deficient in Nox1 (Nox1-KO). Increased nitrotyrosine adduct formation, a marker of peroxynitrite-induced injury, was observed in hepatic sinusoids of WT, which was significantly suppressed in NOX1-KO. NOX1 mRNA was mainly expressed in sinusoidal endothelial cells (LSECs), and it was significantly up-regulated in primary cultured LSECs treated with (PA). The production of nitric oxide by LSECs and LSECs-dependent relaxation of hepatic stellate cells were significantly attenuated by PA treatment. In contrast, these effects of PA were not observed in cells isolated from Nox1-KO. Taken together, the up-regulation of NOX1 in LSECs may elicit peroxynitrite-mediated cellular injury and impair hepatic microcirculation through reduced bioavailability of nitric oxide. ROS derived from NOX1 may therefore constitute a critical component in the development of NASH.

Keyword: fatty liver

miR-192-5p regulates lipid synthesis in non-alcoholic disease through SCD-1.

To evaluate the levels of miR-192-5p in non-alcoholic disease (NAFLD) models and demonstrate the role of miR-192-5p in lipid accumulation.Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet (HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic miR-192-5p and stearoyl-CoA desaturase 1 (SCD-1) levels were measured. MiR-192-5p mimic and inhibitor and SCD-1 siRNA were transfected into Huh7 cells exposed to (PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic miR-192-5p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection ( < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of miR-192-5p and SCD-1 protein levels, respectively ( < 0.01). Transfection with miR-192-5p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively ( < 0.01). Luciferase activity was suppressed and enhanced by miR-192-5p mimic and inhibitor, respectively, in wild-type SCD-1 ( < 0.01) but not in mutant SCD-1. MiR-192-5p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 siRNA transfection abrogated the lipid deposition aggravated by miR-192-5p inhibitor ( < 0.01).This study demonstrates that miR-192-5p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.

Keyword: fatty liver

Hepatocyte-Derived Lipotoxic Extracellular Vesicle Sphingosine 1-Phosphate Induces Macrophage Chemotaxis.

The pathophysiology of non-alcoholic steatohepatitis involves hepatocyte lipotoxicity due to excess saturated free acids and concomitant proinflammatory macrophage effector responses. These include the infiltration of macrophages into hepatic cords in response to incompletely understood stimuli. Stressed hepatocytes release an increased number of extracellular vesicles (EVs), which are known to participate in intercellular signaling and coordination of the behavior of immune cell populations via their cargo. We hypothesized that hepatocyte-derived lipotoxic EVs that are enriched in sphingosine 1-phosphate (S1P) are effectors of macrophage infiltration in the hepatic microenvironment. Lipotoxic EVs were isolated from palmitate treated immortalized mouse hepatocytes and characterized by nanoparticle tracking analysis. Lipotoxic EV sphingolipids were quantified using tandem mass spectrometry. Wildtype and S1P receptor knockout bone marrow-derived macrophages were exposed to lipotoxic EV gradients in a microfluidic gradient generator. Macrophage migration toward EV gradients was captured by time-lapse microscopy and analyzed to determine directional migration. Fluorescence-activated cell sorting along with quantitative PCR and immunohistochemistry were utilized to characterize the cell surface expression of S1P receptor on intrahepatic leukocytes and hepatic expression of S1P receptor, respectively. Palmitate treatment induced the release of EVs. These EVs were enriched in S1P. Palmitate-induced S1P enriched EVs were chemoattractive to macrophages. EV S1P enrichment depended on the activity of sphingosine kinases 1 and 2, such that, pharmacological inhibition of sphingosine kinases 1 and 2 resulted in a significant reduction in EV S1P cargo without affecting the number of EVs released. When exposed to EVs derived from cells treated with palmitate in the presence of a pharmacologic inhibitor of sphingosine kinases 1 and 2, macrophages displayed diminished chemotactic behavior. To determine receptor-ligand specificity, we tested the migration responses of macrophages genetically deleted in the S1P receptor toward lipotoxic EVs. S1P receptor knockout macrophages displayed a marked reduction in their chemotactic responses toward lipotoxic palmitate-induced EVs. Palmitate-induced lipotoxic EVs are enriched in S1P through sphingosine kinases 1 and 2. S1P-enriched EVs activate persistent and directional macrophage chemotaxis mediated by the S1P receptor, a potential signaling axis for macrophage infiltration during hepatic lipotoxicity, and a potential therapeutic target for non-alcoholic steatohepatitis.

Keyword: fatty liver

Macrophage-Specific Hypoxia-Inducible Factor-1α Contributes to Impaired Autophagic Flux in Nonalcoholic Steatohepatitis.

Inflammatory cell activation drives diverse cellular programming during hepatic diseases. Hypoxia-inducible factors (HIFs) have recently been identified as important regulators of immunity and inflammation. In nonalcoholic steatohepatitis (NASH), HIF-1α is upregulated in hepatocytes, where it induces steatosis; however, the role of HIF-1α in macrophages under metabolic stress has not been explored. In this study, we found increased HIF-1α levels in hepatic macrophages in methionine-choline-deficient (MCD) diet-fed mice and in macrophages of patients with NASH compared with controls. The HIF-1α increase was concomitant with elevated levels of autophagy markers BNIP3, Beclin-1, LC3-II, and p62 in both mouse and human macrophages. LysM HIF fl/fl mice, which have HIF-1α levels stabilized in macrophages, showed higher steatosis and inflammation compared with HIF fl/fl mice on MCD diet. In vitro and ex vivo experiments reveal that saturated , (PA), both induces HIF-1α and impairs autophagic flux in macrophages. Using small interfering RNA-mediated knock-down and overexpression of HIF-1α in macrophages, we demonstrated that PA impairs autophagy via HIF-1α. We found that HIF-1α mediates NF-κB activation and MCP-1 production and that HIF-1α-mediated impairment of macrophage autophagy increases IL-1β production, contributing to MCD diet-induced NASH. Conclusion: impairs autophagy via HIF-1α activation in macrophages. HIF-1α and impaired autophagy are present in NASH in vivo in mouse macrophages and in human blood monocytes. We identified that HIF-1α activation and decreased autophagic flux stimulate inflammation in macrophages through upregulation of NF-κB activation. These results suggest that macrophage activation in NASH involves a complex interplay between HIF-1α and autophagy as these pathways promote proinflammatory overactivation in MCD diet-induced NASH.© 2018 by the American Association for the Study of Diseases.

Keyword: fatty liver

Acetone fraction from Sechium edule (Jacq.) S.w. edible roots exhibits anti-endothelial dysfunction activity.

A recent ethnomedical survey on medicinal plants grown in Mexico revealed that Sechium edule (Jacq.) Sw. (Cucurbitaceae) is one of the most valued plant species to treat cardiovascular diseases, including hypertension. Fruits, young leaves, buds, stems, and tuberous roots of the plant are edible. Considering that endothelial dysfunction induced by Angiotensin II plays an important role in the pathogenesis of hypertension and is accompanied by a prooxidative condition, which in turn induces an inflammatory state, vascular remodeling, and tissue damage, and that S. edule has been reported to possess antioxidant, anti-inflammatory and antihypertensive activity, its capability to control endothelial dysfunction was also assessed.To assess in vivo the anti-endothelial dysfunction activity of the acetone fraction (rSe-ACE) of the hydroalcoholic extract from S. edule roots.Endothelial dysfunction was induced in female C57BL/6\u202fJ mice by a daily intraperitoneal injection of angiotensin II for 10 weeks. Either rSe-ACE or losartan (as a control) were co-administered with angiotensin II for the same period. Blood pressure was measured at weeks 0, 5, and 10. Kidney extracts were prepared to determine IL1β, IL4, IL6, IL10, IL17, IFNγ, TNFα, and TGFβ levels by ELISA, along with the prooxidative status as assessed by the activity of antioxidant enzymes. The expression of ICAM-1 was evaluated by immunohistochemistry in kidney histological sections. Kidney and hepatic damage, as well as vascular tissue remodeling, were studied.The rSe-ACE fraction administered at a dose of 10\u202fmg/kg was able to control hypertension, as well as the prooxidative and proinflammatory status in kidney as efficiently as losartan, returning mice to normotensive levels. Additionally, the fraction was more efficient than losartan to prevent and kidney damage. Phytochemical characterization identified cinnamic as a major compound, and linoleic, , and myristic acids as the most abundant non-polar components in the mixture, previously reported to aid in the control of hypertension, inflammation, and oxidative stress, three important components of endothelial dysfunction.this study demonstrated that rSe-ACE has anti-endothelial dysfunction activity in an experimental model and highlights the role of cinnamic and acids in the observed effects.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Acanthoic modulates lipogenesis in nonalcoholic disease via FXR/LXRs-dependent manner.

Acanthoic (AA) is a pimaradiene diterpene isolated from Acanthopanax koreanum Nakai (Araliaceae), with anti-inflammatory and hepatic-protective effects. The present study intended to reveal the effect and mechanism of AA on nonalcoholic disease (NAFLD) associated with lipid accumulation by activating Farnesoid X receptor (FXR) and X receptors (LXRs) signaling. C57BL/6 mice were received a modified Lieber-DeCarli diet with 71% high-fat (L-D) and treated with AA (20 and 40\u202fmg/kg) or equal volume of saline for 12 weeks. The regulation of AA on lipid accumulation was also detected in pro-steatotic stimulated AML12\u202fcells with (PA). When L-D diet-fed mice were treated with AA, loss in body weight, index, and lipid droplet were observed along with reduced triglyceride (TG) and serum transaminase. Furthermore, AA decreased sterol regulatory element binding protein 1 (SREBP-1) and target genes expression, regulated PPARα and PPARγ expressions, ameliorated hepatic fibrosis markers, enhanced hepatic FXR and LXR, and regulated AMPK-LKB1 and SIRT1 signaling pathway. Moreover, AA attenuated lipid accumulation via FXR and LXR activation in steatotic AML-12\u202fcells, which was confirmed by guggulsterones (FXR antagonist) or GW3965 (LXR agonist). Activation of FXR and LXR signaling caused by AA might increase AMPK-SIRT1 signaling and then contribute to modulating lipid accumulation and synthesis, which suggested that activated FXR-LXR axis by AA represented an effective strategy for relieving NAFLD.Copyright © 2019 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Advanced Fibrosis Is Independently Associated with and Insulin Levels in Patients with Non-Alcoholic Disease.

Changes in lipid metabolism occur during the development and progression non-alcoholic disease (NAFLD). However, the (FA) profile in red blood cells (RBC) from patients with fibrosis remains unexplored. Thus, the goal of this study was to evaluate the profile in RBC, dietary lipid intake and insulin resistance indicators in patients with NAFLD, according to the degree of hepatic fibrosis. Using elastography, patients were classified with ( = 52) and without ( = 37) advanced fibrosis. The profile in RBC was analyzed using gas chromatography and the lipid intake was evaluated through a 24-h dietary recall. Subjects with advanced fibrosis had higher levels of , stearic and oleic and total monounsaturated (MUFA) and insulin ( < 0.05), and lower levels of elongase very long chain acids protein-6 and the delta-5-desaturase enzymatic activity ( < 0.05). These results suggest a lack of regulation of enzymes related to FA metabolism in patients with advanced fibrosis.

Keyword: fatty liver

Novel hepatoprotective role of Leonurine hydrochloride against experimental non-alcoholic steatohepatitis mediated via AMPK/SREBP1 signaling pathway.

Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome and is characterized by steatosis, inflammation, and fibrosis. We aim to characterize the hepatoprotective effects of Leonurine hydrochloride (LH) and the possible pathway in a cell and rodent model of diet-induced steatohepatitis (NASH).For in vitro studies, (PA) and free (FFA) induced HepG2 and HL7702 steatosis cell models were used. For in vivo studies, NASH was induced by feeding mice MCD diet. These mice received either placebo or LH at three different doses (50、100、200\u2009mg/kg/day) for 6 weeks. Histological staining\'s, and commercially available kits for ALT and AST and hepatic contents of TG, TC, MDA, SOD, and GSH were used to assess NASH. Furthermore, relative protein and gene expression levels were determined by Western Blot and qPCR, respectively.After establishing NASH models, LH treatment improved lipid accumulation, hepatic contents of TG, TC, and expression levels of ALT and AST in dose-dependent manner. Also, LH improved MDA, SOD, and GSH expression levels. The results of RT-PCR and Western blotting showed that LH upregulated the expression of AMPK phosphorylation and downregulated SREBP-1c and its target genes expression level.Our data reveal the promising role of Leonurine hydrochloride in the prevention and treatment of NASH, in vitro and in vivo. This effect may be partially mediated by the AMPK/SREBP1 pathway. These findings provide a novel therapeutic target for the clinical treatment of NASH.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fatty liver

Euodia daniellii Hemsl. (Bee-Bee Tree) Oil Attenuates Palmitate-Induced Lipid Accumulation and Apoptosis in Hepatocytes.

Hepatic lipid accumulation and apoptosis is elevated in patients with non-alcoholic steatohepatitis and is closely associated with severity. Saturated palmitate stimulates lipid accumulation and apoptosis in hepatocytes. In the present study, we examined bee-bee tree oil (BO)-mediated protective effects on palmitate-induced lipid accumulation and apoptosis in mouse primary hepatocytes. Cells were cultured in a control media or the same media containing 150 or 300 µmol/L of albumin-bound palmitate for 24 h. BO concentrations used were 0, 0.1, 0.2, or 0.5%. Palmitate induced lipid accumulation and mRNA expression of lipogenic genes such as SREBP1c and SCD1. However, BO prevented these changes. Furthermore, palmitate stimulated caspase-3 activity and decreased cell viability in the absence of BO. BO reduced palmitate-induced activation of caspase-3 and cell death in a dose-dependent manner. AMP-activated protein kinase inhibitors abolished the effects of BO. Furthermore, BO suppressed palmitate-induced c-Jun N-terminal kinase (JNK) phosphorylation through the 5' adenosine monophosphate-activated protein kinase (AMPK)-dependent pathway. In conclusion, BO attenuated palmitate-induced hepatic steatosis and apoptosis through AMPK-mediated suppression of JNK signaling. These data suggest that BO is an important determinant of saturated -induced lipid accumulation and apoptosis, and may be an effective therapeutic strategy for treatment of obesity-mediated diseases.© 2018 S. Karger AG, Basel.

Keyword: fatty liver

d- chiro-Inositol Ameliorates High Fat Diet-Induced Hepatic Steatosis and Insulin Resistance via PKCε-PI3K/AKT Pathway.

d- chiro-Inositol (DCI) is a biologically active component found in tartary buckwheat, which can reduce hyperglycemia and ameliorate insulin resistance. However, the mechanism underlying the antidiabetic effects of DCI remains largely unclear. This study investigated the effects and underlying molecular mechanisms of DCI on hepatic gluconeogenesis in mice fed a high fat diet and saturated -treated hepatocytes. DCI attenuated free uptake by the via lipid trafficking inhibition, reduced diacylglycerol deposition, and hepatic PKCε translocation. Thus, DCI could improve insulin sensitivity by suppressing hepatic gluconeogenesis. Subsequent analyses revealed that DCI decreased hepatic glucose output and the expression levels of PEPCK and G6 Pase in insulin resistant mice through PKCε-IRS/PI3K/AKT signaling pathway. Likewise, such effects of DCI were confirmed in HepG2 cells with palmitate-induced insulin resistance. These findings indicate a novel pathway by which DCI prevents hepatic gluconeogenesis, reduces lipid deposition, and ameliorates insulin resistance via regulation of PKCε-PI3K/AKT axis.

Keyword: fatty liver

acids uptake and oxidation are increased in the of rats with adjuvant-induced arthritis.

Severe rheumatoid cachexia is associated with pronounced loss of muscle and fat mass in patients with advanced rheumatoid arthritis. This condition is associated with dyslipidemia and predisposition to cardiovascular diseases. Circulating levels of triglycerides (TG) and free acids (FFA) have not yet been consistently defined in severe arthritis. Similarly, the metabolism of these lipids in the arthritic has not yet been clarified. Aiming at filling these gaps this study presents a characterization of the circulating lipid profile and of the acids uptake and metabolism in perfused livers of rats with adjuvant-induced arthritis. The levels of TG and total cholesterol were reduced in both serum (10-20%) and (20-35%) of arthritic rats. The levels of circulating FFA were 40% higher in arthritic rats, possibly in consequence of cytokine-induced adipose tissue lipolysis. Hepatic uptake and oxidation of and oleic acids was higher in arthritic livers. The phenomenon results possibly from a more oxidized state of the arthritic . Indeed, NADPH/NADP and NADH/NAD ratios were 30% lower in arthritic livers, which additionally presented higher activities of the citric cycle driven by both endogenous and exogenous FFA. The lower levels of circulating and hepatic TG possibly are caused by an increased oxidation associated to a reduced synthesis of acids in arthritic livers. These results reveal that the lipid hepatic metabolism in arthritic rats presents a strong catabolic tendency, a condition that should contribute to the marked cachexia described for arthritic rats and possibly for the severe rheumatoid arthritis.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

PAHSAs enhance hepatic and systemic insulin sensitivity through direct and indirect mechanisms.

esters of hydroxy stearic acids (PAHSAs) are bioactive lipids with antiinflammatory and antidiabetic effects. PAHSAs reduce ambient glycemia and improve glucose tolerance and insulin sensitivity in insulin-resistant aged chow- and high-fat diet-fed (HFD-fed) mice. Here, we aimed to determine the mechanisms by which PAHSAs improve insulin sensitivity. Both acute and chronic PAHSA treatment enhanced the action of insulin to suppress endogenous glucose production (EGP) in chow- and HFD-fed mice. Moreover, chronic PAHSA treatment augmented insulin-stimulated glucose uptake in glycolytic muscle and heart in HFD-fed mice. The mechanisms by which PAHSAs enhanced hepatic insulin sensitivity included direct and indirect actions involving intertissue communication between adipose tissue and . PAHSAs inhibited lipolysis directly in WAT explants and enhanced the action of insulin to suppress lipolysis during the clamp in vivo. Preventing the reduction of free acids during the clamp with Intralipid infusion reduced PAHSAs\' effects on EGP in HFD-fed mice but not in chow-fed mice. Direct hepatic actions of PAHSAs may also be important, as PAHSAs inhibited basal and glucagon-stimulated EGP directly in isolated hepatocytes through a cAMP-dependent pathway involving Gαi protein-coupled receptors. Thus, this study advances our understanding of PAHSA biology and the physiologic mechanisms by which PAHSAs exert beneficial metabolic effects.

Keyword: fatty liver

Wolf Extract Ameliorates Hepatic Steatosis through Regulation of Lipid Metabolism, Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation.

Wolf (PCW) is an edible, pharmaceutical mushroom with remarkable biological properties including anti-tumor, anti-inflammation, anti-oxidation, anti-ageing, and anti-diabetic effects. In the current study, we investigated the effects of PCW extract on hepatic steatosis under in vitro and in vivo conditions, and elucidated the underlying mechanisms. In this study, a mixture of HepG2 cells treated with free (FFA)- and oleic -and high-fat diet (HFD)-fed obese mice were used; in this background, the triglyceride (TG) levels in HepG2 cells and mice were measured, and the expression levels of genes associated with lipogenesis, oxidation, endoplasmic reticulum (ER) stress, and autophagy were determined. Treatment of HepG2 cells with FFA enhanced intracellular TG levels in HepG2 cells, but co-treatment with PCW significantly attenuated the TG levels. Notably, PCW significantly enhanced the phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein-1c (SREBP-1c) in FFA-treated HepG2 cells. PCW downregulated the expression of lipogenesis-related genes, but upregulated the expression of genes associated with oxidation. Further, PCW inhibited FFA-induced expression of ER stress markers and induced autophagy proteins. However, inhibition of AMPK significantly attenuated the beneficial effects of PCW in HepG2 cells. Moreover, PCW efficiently decreased HFD-induced hepatic TG accumulation in vivo and increased the phosphorylation of hepatic AMPK. Three compounds present in PCW including poricoic , pachymic , and ergosterol, significantly decreased FFA-induced increase in intracellular TG levels, consistent with increased AMPK phosphorylation, suggesting that poricoic , pachymic , and ergosterol are responsible for PCW-mediated amelioration of hepatic steatosis. Taken together, these results demonstrated that PCW ameliorates hepatic steatosis through the regulation of lipid metabolism, inhibition of ER stress, and activation of autophagy in an AMPK-dependent manner. This suggested that PCW can be potentially used for the treatment of hepatic steatosis.

Keyword: fatty liver

7-Hydroxymatairesinol improves body weight, fat and sugar metabolism in C57BJ/6 mice on a high-fat diet.

7-Hydroxymatairesinol (7-HMR) is a plant lignan abundant in various concentrations in plant foods. The objective of this study was to test HMRLignan™, a purified form of 7-HMR, and the corresponding Picea abies extract (total extract P. abies; TEP) as dietary supplements on a background of a high-fat diet (HFD)-induced metabolic syndrome in mice and in the 3T3-L1 adipogenesis model. Mice, 3 weeks old, were fed a HFD for 60 d. Subgroups were treated with 3 mg/kg body weight 7-HMR (HMRLignan™) or 10 mg/kg body weight TEP by oral administration. 7-HMR and TEP limited the increase in body weight (-11 and -13 %) and fat mass (-11 and -18 %) in the HFD-fed mice. Epididymal adipocytes were 19 and -12 % smaller and the was less steatotic (-62 and -65 %). Serum lipids decreased in TEP-treated mice (-11 % cholesterol, -23 % LDL and -15 % TAG) and sugar metabolism was ameliorated by both lignan preparations, as shown by a more than 70 % decrease in insulin secretion and insulin resistance. The expression of several metabolic genes was modulated by the HFD with an effect that was reversed by lignan. In 3T3-L1 cells, the 7-HMR metabolites enterolactone (ENL) and enterodiol (END) showed a 40 % inhibition of cell differentiation accompanied by the inhibited expression of the adipogenic genes PPARγ, C/EBPα and aP2. Furthermore, END and ENL caused a 10 % reduction in TAG uptake in HEPA 1-6 hepatoma cells. In conclusion, 7-HMR and TEP reduce metabolic imbalances typical of the metabolic syndrome and obesity in male mice, whereas their metabolites inhibit adipogenesis and lipid uptake in vitro.

Keyword: fatty liver

ester of tetrahydrocannabinol (THC) and diester of 11-hydroxy-THC - Unsuccessful search for additional THC metabolites in human body fluids and tissues.

conjugates of hydroxy-metabolites of tetrahydrocannabinol (THC) or cannabinol have already been reported as metabolites in rats. In the herein presented investigation, esters of THC and its primary metabolite 11-hydroxy-delta9-tetrahydrocannabinol (11-OH-THC) were synthesized using esterification with chloride. Structural elucidation of the products was conducted using nuclear magnetic resonance spectroscopy (NMR) and liquid chromatography coupled to quadrupole time of flight mass spectrometry (LC-QToF-MS). For the confirmation of a previous cannabis use, body fluids (femoral blood, heart blood, urine, bile) of 27 death cases (all with known cannabis use), including adipose tissue homogenates of six of these cases as well as eleven plasma samples (probably all with regular cannabis use, confirmed by a high 11-nor-9-carboxy-delta9-tetrahydrocannabinol (THC-COOH) concentration (except one sample, >200ng/mL), were tested for THC and its main metabolites 11-OH-THC and THC-COOH using gas chromatography coupled to mass spectrometry (GC-MS). These samples as well as further tissue homogenates of autopsy cases (, kidney, brain) were additionally tested for the presence of THC ester or 11-OH-THC diester by means of a liquid chromatographic triple quadrupole mass spectrometric (LC-QQQ-MS) method, in order to evaluate a possible presence of these conjugates in humans. In none of the analyzed samples (in total 196 specimens; plasma (N=11), femoral blood (N=23), heart blood (N=25), urine (N=23), bile (N=27), (N=27), kidney (N=27), brain (N=27), adipose tissue (N=6)), esters of THC or 11-OH-THC could be proven. Even if the existence of these esters in human samples cannot be ruled out definitely, suitability as cannabis consumption markers does not seem likely based on our findings.Copyright © 2018 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Treatment of cigarette smoke extract and condensate differentially potentiates -induced lipotoxicity and steatohepatitis in vitro.

Accumulative evidence showed that cigarette smoke (CS) detrimentally affects the pathogenesis of nonalcoholic steatohepatitis (NASH). The purpose of this study was to evaluate the effects of CS extract (CSE) or total particulate matter (TPM) on the in vitro steatohepatitis model using mouse primary hepatocytes treated with (PA) or PA plus LPS. Increased hepatocellular damage was observed in PA-treated hepatocytes with TPM or CSE treatment, but increased triglyceride level was only observed in PA plus LPS-treated hepatocytes with a high concentration of TPM. Also, expression levels of steatohepatitis-related genes such as TNF-α, NOS 2, and SREBP-1c were significantly increased after treatment of TPM. To further demonstrate the role of Kupffer cells (KCs) after CS extracts treatment, trans-well co-culture system of hepatocytes and KCs was utilized. The levels of inflammatory cytokines and the ratios of Bax/Bcl-2 (apoptosis-related genes) were markedly increased in co-cultured hepatocytes after TPM or CSE treatment. Interestingly, KCs activation was augmented in KCs upon treatment with CSE or TPM. Overall, our findings indicate that in vitro treatment with CSE or TPM differentially contributes to the severity of steatohepatitis by modulating steatohepatitis-related lipotoxicity and inflammation, which might be caused by KCs activation with subsequent induction of hepatocytes apoptosis.Copyright © 2018 Elsevier Ltd. All rights reserved.

Keyword: fatty liver

Cathepsin B inhibition ameliorates the non-alcoholic steatohepatitis through suppressing caspase-1 activation.

Non-alcoholic disease (NAFLD) has emerged as the most common chronic disease. NLRP3 inflammasome activation has been widely studied in the pathogenesis of NAFLD. Cathepsin B (CTSB) is a ubiquitous cysteine cathepsin, and the role of CTSB in the progression and development of NAFLD has received extensive concern. However, the exact roles of CTSB in the NAFLD development and NLRP3 inflammasome activation are yet to be evaluated. In the present study, we used methionine choline-deficient (MCD) diet to establish mice NASH model. CTSB inhibitor (CA-074) was used to suppress the expression of CSTB. Expressions of CTSB and caspase-1 were evaluated by immunohistochemical staining. Serum IL-1β and IL-18 levels were also determined. was used to stimulate Kupffer cells (KCs), and protein expressions of CTSB, NLRP3, ASC (apoptosis-associated speck-like protein containing CARD), and caspase-1 in KCs were detected. The levels of IL-1β and IL-18 in the supernatant of KCs were evaluated by enzyme-linked immunosorbent assay (ELISA). Our results showed that CTSB inhibition improved the function and reduced hepatic inflammation and ballooning, and the levels of pro-inflammatory cytokines IL-1β and IL-18 were decreased. The expressions of CTSB and caspase-1 in tissues were increased in the NASH group. In in vitro experiments, PA stimulation could increase the expressions of CTSB and NLRP3 inflammasome in KCs, and CTSB inhibition downregulated the expression of NLRP3 inflammasome in KCs, when challenged by PA. Moreover, CTSB inhibition effectively suppressed the expression and activity of caspase-1 and subsequently secretions of IL-1β and IL-18. Collectively, these results suggest that CTSB inhibition limits NLRP3 inflammasome-dependent NASH formation through regulating the expression and activity of caspase-1, thus providing a novel anti-inflammatory signal pathway for the therapy of NAFLD.

Keyword: fatty liver

Unsaturated Acids Increase the Expression of Hepassocin through a Signal Transducer and Activator of Transcription 3-Dependent Pathway in HepG2 Cells.

Hepassocin (HPS) is a hepatokine that regulates hepatocyte proliferation. It is known that HPS plays an important role in the development of nonalcoholic diseases (NAFLD). acids, such as oleic (OLA), exhibit the ability to activate the signal transducer and activator of transcription-3 (STAT3), and the binding site of STAT3 is found in the promoter region of HPS. However, the regulation of HPS by acids is still obscure. To clarify the regulation of HPS, we detected the expression of HPS by western blots. In addition, a hepatic steatosis cell culture model was established by treatment of different acids, including linoleic (LNA), oleic , , and stearic . The intracellular lipid accumulation was confirmed by oil red O staining. Blocking of STAT3 activity was achieved by the pretreatment of the STAT3 inhibitor, stattic. We found that activation of STAT3 by interleukin-6 (IL-6) was mediated in the regulation of HPS expression. Treatment of unsaturated acids significantly induced intracellular lipid accumulation in HepG2 cells. Moreover, the expressions of HPS were increased in unsaturated -treated HepG2 cells, as compared with saturated -treated groups. Also, the expression of HPS induced by OLA was blocked by the inhibition of STAT3 activity. Furthermore, we found that deletion of HPS by small interfering ribonucleic transfection decreased the protective effect of OLA on cell viability. Taken together, we provided evidence that STAT3 plays an important role in the regulation of OLA-induced HPS expression and the increased HPS may further participate in the development of NAFLD. In addition, the increase of HPS might be involved in the protective effect of OLA on cell viability.© 2018 AOCS.

Keyword: fatty liver

Degradation of splicing factor SRSF3 contributes to progressive liver disease.

Serine rich splicing factor 3 (SRSF3) plays a critical role in liver function and its loss promotes chronic liver damage and regeneration. As a consequence, genetic deletion of SRSF3 in hepatocytes caused progressive liver disease and ultimately led to hepatocellular carcinoma. Here we show that SRSF3 is decreased in human liver samples with non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or cirrhosis that was associated with alterations in RNA splicing of known SRSF3 target genes. Hepatic SRSF3 expression was similarly decreased and RNA splicing dysregulated in mouse models of NAFLD and NASH. We showed that -induced oxidative stress caused conjugation of the ubiquitin like NEDD8 protein to SRSF3 and proteasome mediated degradation. SRSF3 was selectively neddylated at lysine11 and mutation of this residue (SRSF3-K11R) was sufficient to prevent both SRSF3 degradation and alterations in RNA splicing. Finally prevention of SRSF3 degradation in vivo partially protected mice from hepatic steatosis, fibrosis and inflammation. These results highlight a neddylation-dependent mechanism regulating gene expression in the liver that is disrupted in early metabolic liver disease and may contribute to the progression to NASH, cirrhosis and ultimately hepatocellular carcinoma.

Keyword: fatty liver

Dihydrosterculic from cottonseed oil suppresses desaturase activity and improves metabolomic profiles of high-fat-fed mice.

Polyunsaturated (PUFA)-rich diets are thought to provide beneficial effects toward metabolic health in part through their bioactive properties. We hypothesized that increasing PUFA intake in mice would increase peroxisome proliferator activated receptor delta (PPARδ) expression and activity, and we sought to examine the effect of different PUFA-enriched oils on muscle PPARδ expression. One of the oils we tested was cottonseed oil (CSO) which is primarily linoleic (53%) and (24%). Mice fed a CSO-enriched diet (50% energy from fat) displayed no change in muscle PPARδ expression; however, in the , it was consistently elevated along with its transcriptional coactivator Pgc-1. Male mice were fed chow or CSO-, saturated fat (SFA)-, or linoleic (18:2)-enriched diets that were matched for macronutrient content for 4 weeks. There were no differences in food intake, body weight, fasting glucose, glucose tolerance, or energy expenditure between chow- and CSO-fed mice, whereas SFA-fed mice had increased fat mass and 18:2-fed mice were less glucose tolerant. Metabolomic analyses revealed that the livers of CSO-fed mice closely matched those of chow-fed but significantly differed from SFA- and 18:2-enriched groups. composition of the diets and livers revealed an impairment in desaturase activity and the presence of dihydrosterculic (DHSA) in the CSO-fed mice. The effect of DHSA on PPARδ and stearoyl-CoA desaturase-1 expression mimicked that of the CSO-fed mice. Taken together, these data suggest that DHSA from CSO may be an effective means to increase PPARδ expression with concomitant suppression of stearoyl-CoA desaturase-1 activity.Copyright © 2017 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Water Extract of L. Ameliorates Non-Alcoholic Disease.

Our aim was to investigate whether hot water extract (CLW) of L. could prevent non-alcoholic disease (NAFLD). HepG2 cells were treated with free (FFA) mixture (oleic : , 2:1) for 24 h to stimulate in vitro . In addition, C57BL/6 mice were fed 60 kcal% high-fat (HF) diet for eight weeks to induce in vivo. Intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) productions were increased by FFA and HF-diet, but supplementation with CLW significantly decreased these levels. CLW treatment ameliorated antioxidant activities that were suppressed by exposure to the FFA and HF-diet. Cluster of differentiation 36 (CD36) and transport proteins (FATP2 and FATP5) were increased in HF-diet groups, while CLW suppressed their expression levels. Moreover, sterol regulatory element-binding protein-1c (SREBP-1c), acetyl-coenzyme A carboxylase (ACC), and synthase (FAS) expression levels were down-regulated in the CLW groups compared to HF-diet groups. On the other hand, 5\' adenosine monophosphate-activated protein kinase (AMPK), Peroxisome proliferator-activated receptor alpha (PPAR-α), and carnitine palmitoyltransferase 1 (CPT-1) expressions were up-regulated in the CLW groups. HF-diet fed mice showed high hepatic triglycerides (TG) content compared to the normal diet mice. However, the administration of CLW restored the hepatic TG level, indicating an inhibitory effect against lipid accumulation by CLW. These results suggest that CLW could be a potentially useful agent for the prevention of NAFLD through modulating uptake.

Keyword: fatty liver

Gut microbiota-mediated generation of saturated acids elicits inflammation in the in murine high-fat diet-induced steatohepatitis.

The gut microbiota plays crucial roles in the development of non-alcoholic steatohepatitis (NASH). However, the precise mechanisms by which alterations of the gut microbiota and its metabolism contributing to the pathogenesis of NASH are not yet fully elucidated.Mice were fed with a recently reported new class of high-fat diet (HFD), steatohepatitis-inducing HFD (STHD)-01 for 9\xa0weeks. The composition of the gut microbiota was analyzed by T-RFLP. Luminal metabolome was analyzed using capillary electrophoresis and liquid chromatography time-of-flight mass spectrometry (CE- and LC-TOFMS).Mice fed the STHD-01 developed NASH-like pathology within a short period. Treatment with antibiotics prevented the development of NASH by STHD-01. The composition of the gut microbiota and its metabolic activities were markedly perturbed in the STHD-01-fed mice, and antibiotic administration normalized these changes. We identified that long-chain saturated and n-6 metabolic pathways were significantly altered by STHD-01. Of note, the changes in gut lipidome caused by STHD-01 were mediated by gut microbiota, as the depletion of the gut microbiota could reverse the perturbation of these metabolic pathways. A saturated long-chain , , which accumulated in the STHD-01 group, activated macrophages and promoted TNF-α expression.Lipid metabolism by the gut microbiota, particularly the saturation of acids, affects fat accumulation in the and subsequent inflammation in NASH.

Keyword: fatty liver

Evidence for an alternative desaturation pathway increasing cancer plasticity.

Most tumours have an aberrantly activated lipid metabolism that enables them to synthesize, elongate and desaturate acids to support proliferation. However, only particular subsets of cancer cells are sensitive to approaches that target metabolism and, in particular, desaturation. This suggests that many cancer cells contain an unexplored plasticity in their metabolism. Here we show that some cancer cells can exploit an alternative desaturation pathway. We identify various cancer cell lines, mouse hepatocellular carcinomas, and primary human and lung carcinomas that desaturate palmitate to the unusual sapienate to support membrane biosynthesis during proliferation. Accordingly, we found that sapienate biosynthesis enables cancer cells to bypass the known desaturation pathway that is dependent on stearoyl-CoA desaturase. Thus, only by targeting both desaturation pathways is the in vitro and in vivo proliferation of cancer cells that synthesize sapienate impaired. Our discovery explains metabolic plasticity in desaturation and constitutes an unexplored metabolic rewiring in cancers.

Keyword: fatty liver

Inhibition of MD2-dependent inflammation attenuates the progression of non-alcoholic disease.

Non-alcoholic disease (NAFLD) can progress to the more serious non-alcoholic steatohepatitis (NASH), characterized by inflammatory injury and fibrosis. The pathogenic basis of NAFLD progressing to NASH is currently unknown, but growing evidence suggests MD2 (myeloid differentiation factor 2), an accessory protein of TLR4, is an important signalling component contributing to this disease. We evaluated the effectiveness of the specific MD2 inhibitor, L6H21, in reducing inflammatory injury in a relevant high-fat diet (HFD) mouse model of NASH and in the (PA)-stimulated human cell line (HepG2). For study, genetic knockout (MD2 ) mice were fed a HFD or control diet for 24 weeks, or wild-type mice placed on a similar diet regimen and treated with L6H21 for the last 8 or 16 weeks. Results indicated that MD2 inhibition with L6H21 was as effective as MD2 knockout in preventing the HFD-induced hepatic lipid accumulation, pro-fibrotic changes and expression of pro-inflammatory molecules. Direct challenge of HepG2 with PA (200 μM) increased MD2-TLR4 complex formation and expression of pro-inflammatory and pro-fibrotic genes and L6H21 pre-treatment prevented these PA-induced responses. Interestingly, MD2 knockout or L6H21 increased expression of the anti-inflammatory molecule, PPARγ, in tissue and the cell line. Our results provide further evidence for the critical role of MD2 in the development of NASH and conclude that MD2 could be a potential therapeutic target for NAFLD/NASH treatment. Moreover, the small molecule MD2 inhibitor, L6H21, was an effective and selective investigative agent for future mechanistic studies of MD2.© 2017 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fatty liver

Liraglutide protects non-alcoholic disease via inhibiting NLRP3 inflammasome activation in a mouse model induced by high-fat diet.

Liraglutide, a glucagon-like peptide-1 (GLP-1) analogue that has recently become the first-line treatment for type 2 diabetes mellitus (T2DM), has also been reported to decrease degeneration of the . The purpose of this study is to explore whether liraglutide improves high-fat diet-induced non-alcoholic disease (NAFLD) in mice through inhibiting the NLRP3 inflammasome in the . After daily intraperitoneal injection of liraglutide (0.6\u202fmg/kg body weight) for four weeks, the , /body weight, serum levels of ALT, AST, total cholesterol, triglycerides and LDL were significantly decreased in a high-fat diet-induced NAFLD mouse model. The hepatic steatosis among sections of H&E and Oil Red O staining was also markedly reduced after treatment with liraglutide. The expressions of NLRP3 inflammasome components (including NLRP3, ASC, and caspase-1) in the of mice after treatment with liraglutide were decreased substantially. In\xa0vitro studies found that the mitochondrial dysfunction in Kupffer cells induced by was attenuated, and the protein levels of NLRP3, ASC and caspase-1 were also decrease markedly. These results demonstrate that liraglutide was able to alleviate high-fat diet-induced hepatic steatosis via inhibiting NLRP3 inflammasome activation, suggesting that liraglutide is a potent drug that can reverse the pathological hallmarks of NAFLD.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Impact of novel palmitoylated prolactin-releasing peptide analogs on metabolic changes in mice with diet-induced obesity.

Analogs of anorexigenic neuropeptides, such as prolactin-releasing peptide (PrRP), have a potential as new anti-obesity drugs. In our previous study, attached to the N-terminus of PrRP enabled its central anorexigenic effects after peripheral administration. In this study, two linkers, γ-glutamic at Lys11 and a short, modified polyethylene glycol at the N-terminal Ser and/or Lys11, were applied for the palmitoylation of PrRP31 to improve its bioavailability. These analogs had a high affinity and activation ability to the PrRP receptor GPR10 and the neuropeptide FF2 receptor, as well as short-term anorexigenic effect similar to PrRP palmitoylated at the N-terminus. Two-week treatment with analogs that were palmitoylated through linkers to Lys11 (analogs 1 and 2), but not with analog modified both at the N-terminus and Lys11 (analog 3) decreased body and weights, insulin, leptin, triglyceride, cholesterol and free plasma levels in a mouse model of diet-induced obesity. Moreover, the expression of uncoupling protein-1 was increased in brown fat suggesting an increase in energy expenditure. In addition, treatment with analogs 1 and 2 but not analog 3 significantly decreased urinary concentrations of 1-methylnicotinamide and its oxidation products N-methyl-2-pyridone-5-carboxamide and N-methyl-4-pyridone-3-carboxamide, as shown by NMR-based metabolomics. This observation confirmed the previously reported increase in nicotinamide derivatives in obesity and type 2 diabetes mellitus and the effectiveness of analogs 1 and 2 in the treatment of these disorders.

Keyword: fatty liver

Trimethylamine N-Oxide Aggravates Steatosis through Modulation of Bile Metabolism and Inhibition of Farnesoid X Receptor Signaling in Nonalcoholic Disease.

Trimethylamine N-oxide (TMAO), the metabolite of choline generated by gut microbiota, is associated with nonalcoholic disease (NAFLD) and could influence bile (BA) metabolism. However, whether TMAO aggravates steatosis by modulating BA metabolism and the related mechanisms has not been investigated.A case-control study including biopsy-proven NAFLD patients (n\xa0=\xa034) and controls (n\xa0=\xa014) is conducted to determine the correlation between TMAO and BA metabolism. Serum levels of total BA and the percentage of farnesoid X receptor (FXR)-antagonistic BA species are markedly higher in NAFLD patients than in the controls. Serum levels of TMAO positively correlated with the serum levels of total BA and hepatic mRNA expression of cholesterol 7 alpha hydroxylase (CYP7A1). In a murine model, it is found that 18 weeks administration of TMAO impairs function and increases hepatic triglyceride accumulation and lipogenesis in mice fed with a high-fat diet. TMAO increases BA synthesis and shifted hepatic BA composition toward FXR-antagonistic activity. Knockdown of CYP7A1 via small interfering RNA or activation of FXR by GW4064 blocks the effect of TMAO-induced lipogenesis in -treated HepG2 cells.TMAO aggravates steatosis by suppressing BA-mediated hepatic FXR signaling.© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fatty liver

Natural Abundance Carbon Isotopic Analysis Indicates the Equal Contribution of Local Synthesis and Plasma Uptake to Palmitate Levels in the Mouse Brain.

Saturated acids are the most abundant acids in the brain, however, there has been some debate regarding the ability of intact dietary saturated acids to be incorporated into the brain. In the present study, we use compound specific isotope analysis to measure the natural abundance carbon isotopic signature of brain, , and blood (PAM) and compare it to the dietary PAM and sugar isotopic signatures to calculate the relative contribution of both the incorporation of intact and endogenously synthesized PAM to these pools. Mice were equilibrated to the study diet, and extracted acids were analyzed with gas chromatography isotope ratio mass spectrometry to determine the carbon isotopic signature of PAM (δ C ). , serum total, and serum unesterified δ C ranged between -20.6 and -21.1 mUr and were approximately 8.5 mUr more enriched in C when compared to the dietary PAM signature. Brain δ C was found to be more enriched than or blood pools (-16.7 ± 0.2 mUr, mean ± SD). Two end-member-mixed modeling using the carbon isotopic signature of dietary PAM and dietary sugars determined the contribution of synthesis to the total tissue PAM pool to range between 44% and 48%. This suggests that endogenous synthesis and dietary PAM are near equal contributors to brain, , and blood PAM pools. In conclusion, our data provide evidence that brain PAM levels are maintained by both local endogenous synthesis and through the uptake of intact PAM from the blood.© 2018 AOCS.

Keyword: fatty liver

Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of diabetes, obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated in vitro. HFD-induced inflammatory response and function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 conditional knockout mice (miR-429), fed either a normal diet or an HFD, showed severe inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Keyword: fatty liver

Procyanidin B2 ameliorates free acids-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state.

Procyanidin B2, a naturally occurring phenolic compound, has been reported to exert multiple beneficial functions. However, the effect of procyanidin B2 on free acids (FFAs)-induced hepatic steatosis remains obscure. The present study is therefore aimed to elucidate the protective effect of procyanidin B2 against hepatic steatosis and its underlying mechanism. Herein, we reported that procyanidin B2 attenuated FFAs-induced lipid accumulation and its associated oxidative stress by scavenging excessive ROS and superoxide anion radicals, blocking loss of mitochondrial membrane potential, restoring glutathione content, and increasing activity of antioxidant enzymes (GPx, SOD and CAT) in hepatocytes. Procyanidin B2 mechanistically promoted lipid degradation via modulation of transcription factor EB (TFEB), a master regulator of lysosomal pathway. Molecular docking analysis indicated a possible ligand-binding position of procyanidin B2 with TFEB. In addition, administration of procyanidin B2 resulted in a significant reduction of hepatic fat accumulation in high-fat diet (HFD)-induced obese mice, and also ameliorated HFD-induced metabolic abnormalities, including hyperlipidemia and hyperglycemia. It was confirmed that procyanidin B2 prevented HFD-induced hepatic fat accumulation through down-regulating lipogenesis-related gene expressions (PPARγ, C/EBPα and SREBP-1c), inhibiting pro-inflammatory cytokines production (IL-6 and TNF-α) and increasing antioxidant enzymes activity (GPx, SOD and CAT). Moreover, hepatic acids analysis indicated that procyanidin B2 caused a significant increase in the levels of , oleic and linoleic . Intriguingly, procyanidin B2 restored the decreased nuclear TFEB expression in HFD-induced steatosis and up-regulated its target genes involved in lysosomal pathway (Lamp1, Mcoln, Uvrag), which suggested a previously unrecognized mechanism of procyanidin B2 on ameliorating HFD-induced hepatic steatosis. Taken together, our results demonstrated that procyanidin B2 attenuated FFAs-induced hepatic steatosis through regulating TFEB-mediated lysosomal pathway and redox state, which had important implications that modulation of TFEB might be a potential therapeutic strategy for hepatic steatosis and procyanidin B2 could represent a promising novel agent in the prevention and treatment of non-alcoholic disease (NAFLD).Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

A novel recombinant peptide INSR-IgG4Fc (Yiminsu) restores insulin sensitivity in experimental insulin resistance models.

Type 2 diabetes mellitus (T2DM) is a chronic degenerative endocrine and metabolic disease with high mortality and morbidity, yet lacks effective therapeutics. We recently generated a novel fusion peptide INSR-IgG4Fc, Yiminsu (YMS), to facilitate the high-affinity binding and transportation of insulin. Thus, the aim of the present study was to determine whether the novel recombinant peptide, YMS, could contribute to restoring insulin sensitivity and glycaemic control in insulin resistance models and revealing its underlying mechanism. (PA)-treated LO2 cells and high fat diet (HFD)-fed mice were treated with YMS. Therapeutic effects of YMS were measured using Western blotting, ELISA, qPCR, Histology and transmission electron microscopy. We observed that YMS treatment effectively improved insulin signaling in PA-treated LO2 cells and HFD-fed mice. Notably, YMS could significantly reduce serum levels of glucose, triglycerides, acids and cholesterol without affecting the serum insulin levels. Moreover, our data demonstrated that YMS could restore glucose and lipid homeostasis via facilitating insulin transportation and reactivating PI3K/Akt signaling in both PA-treated cells and , gastrocnemius and brown fat of HFD-fed mice. Additionally, we noticed that the therapeutic effects of YMS was similar as rosiglitazone, a well-recognized insulin sensitizer. Our findings suggested that YMS is a potentially candidate for pharmacotherapy for metabolic disorders associated with insulin resistance, particularly in T2DM.Copyright © 2018. Published by Elsevier Masson SAS.

Keyword: fatty liver

[Glucagon-like peptide-1 regulates lipid metabolism in hepatocytes through Foxo1/3].

Glucagon-like peptide-1 (GLP-1) has been reported to be effective in the treatment of nonalcoholic disease (NAFLD). However, the molecular mechanism of GLP-1 on NAFLD is remained unclear. The present study was to detect whether the effect of GLP-1 on triglyceride (TG) content in hepatocytes is dependent on Foxos. HepG2 cells were treated with /oleic for 24 h. The knockdown of Foxo1, Foxo3 was conducted through small interfering RNA (siRNA). Real time PCT (RT-PCR) was used to detect the changes of the SREBP1c and Aco genes in HepG2 cells after Foxo1/3 knockdown. As expected, /oleic increased TG concentration in HepG2 cells [(12.65 ± 1.32) μg/mg vs. (4.32±0.54) μg/mg, 0.05]. Addition of GLP-1 dose (10, 50, 100nmol/L) dependently lowered the TG content and reached plateau at 100 nmol/L of GLP-1 [TG(8.38±1.47) μg/mg]. The GLP-1 effect on TG remained after knocking down either Foxo1 [(9.09±1.34)μg/mg] or Foxo3 [(8.90±1.60) μg/mg] alone, but not when knocking down Foxo1 and Foxo3 (Foxo1/3) together [(14.66±1.77)μg/mg]. Moreover, knocking down Foxo1/3 also abolished GLP-1 effect on SREBP1c and Aco expression. GLP-1 can inhibit the synthesis of TG in hepatocytes depending on Foxo1 and Foxo3. Further studies are needed to explore the specific mechanisms.

Keyword: fatty liver

Sodium tanshinone IIA sulfonate ameliorates hepatic steatosis by inhibiting lipogenesis and inflammation.

Non-alcoholic disease (NAFLD) is becoming an epidemic disease in adults and children worldwide. Importantly, there are currently no approved treatments available for NAFLD. This study aims to investigate the potential applications of sodium tanshinone IIA sulfonate (STS) on improving the NAFLD condition using both in vitro and in vivo approaches. The results showed that STS markedly inhibited lipid accumulation in oleic (OA) and (PA) treated HepG2 and primary immortalized human hepatic (PIH) cells. STS suppressed lipogenesis by inhibiting expression of sterol regulatory element binding transcription factor 1 (SREBF1), synthase (FASN) and stearoyl-CoA desaturase (SCD). In addition, STS reduced inflammation in cells treated with OA-PA, shown by decreased transcriptional levels of tumor necrosis factor (TNF), transforming growth factor beta 1 (TGFB1) and interleukin 1 beta (IL1B). Consistently, protective effects on hepatic steatosis in db/db mice were observed after STS administration, demonstrated by decreased lipid accumulation in mouse hepatocytes. This protective effect might be associated with STS induced activation of sirtuin 1 (SIRT1)/protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1) pathways. Our findings suggest a potential therapeutic role for STS in the treatment of NAFLD.Copyright © 2018 The Authors. Published by Elsevier Masson SAS.. All rights reserved.

Keyword: fatty liver

Resveratrol Maintains Lipid Metabolism Homeostasis via One of the Mechanisms Associated with the Key Circadian Regulator Bmal1.

Resveratrol (RES) possesses anti-inflammatory and anti-oxidant activities, and it can prevent liver lipid metabolism disorders in obese and diabetic individuals. This study elucidated the mechanisms of brain and muscle Arnt-like protein-1 (Bmal1) in the protective effects of RES against liver lipid metabolism disorders. The results indicated that RES ameliorated free fatty (FFA)-induced (oleic (OA): (PA) = 2:1) glycolipid metabolic disorders in hepatocytes. Simultaneously, RES partially reverted the relatively shallow daily oscillations of FFA-induced circadian clock gene transcription and protein expression in HepG2 cells. RES also attenuated FFA-triggered reactive oxygen species (ROS) secretion and restored mitochondrial membrane potential consumption, as well as the restoration of mitochondrial respiratory complex expression. This study provides compelling evidence that RES controls intracellular lipid metabolic imbalance in a Bmal1-dependent manner. Overall, RES may serve as a promising natural nutraceutical for the regulation of lipid metabolic disorders relevant to the circadian clock.

Keyword: fatty liver

Diosgenin ameliorates -induced lipid accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.

Non-alcoholic disease (NAFLD) is the most common chronic disease and is characterized by excessive hepatic lipid accumulation. Many studies have suggested that lipid overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on energy metabolism, but the underlying mechanism remains unclear.Human normal hepatocytes (LO2 cells) were incubated with to establish the cell model of nonalcoholic . The effects of DSG on lipid metabolism, glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, lipid consumption and lipid synthesis in LO2 cells was investigated.The results indicated that induced obvious lipid accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular lipid content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of lipid synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group.This study suggests that DSG can reduce intracellular lipid accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing β-oxidation and decreasing lipid synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.

Keyword: fatty liver

Serine prevented high-fat diet-induced oxidative stress by activating AMPK and epigenetically modulating the expression of glutathione synthesis-related genes.

Serine deficiency has been observed in patients with nonalcoholic disease (NAFLD). Whether serine supplementation has any beneficial effects on the prevention of NAFLD remains unknown. The present study was conducted to investigate the effects of serine supplementation on hepatic oxidative stress and steatosis and its related mechanisms. Forty male C57BL/6J mice (9week-old) were randomly assigned into four groups (n=10) and fed: i) a low-fat diet; ii) a low-fat diet supplemented with 1% (wt:vol) serine; iii) a high-fat (HF) diet; and iv) a HF diet supplemented with 1% serine, respectively. (PA)-treated primary hepatocytes separated from adult mice were also used to study the effects of serine on oxidative stress. The results showed that serine supplementation increased glucose tolerance and insulin sensitivity, and protected mice from hepatic lipid accumulation, but did not significantly decreased HF diet-induced weight gain. In addition, serine supplementation protected glutathione (GSH) antioxidant system and prevented hypermethylation in the promoters of glutathione synthesis-related genes, while decreasing reactive oxygen species (ROS) in mice fed a HF diet. Moreover, we found that serine supplementation increased phosphorylation and S-glutathionylation of AMP-activated protein kinase α subunit (AMPKα), and decreased ROS, malondialdehyde and triglyceride contents in PA-treated primary hepatocytes. However, while AMPK activity or GSH synthesis was inhibited, the abovementioned effects of serine on PA-treated primary hepatocytes were not observed. Our results suggest that serine supplementation could prevent HF diet-induced oxidative stress and steatosis by epigenetically modulating the expression of glutathione synthesis-related genes and through AMPK activation.Copyright © 2017 Elsevier B.V. All rights reserved.

Keyword: fatty liver

Acetyl-CoA from inflammation-induced acids oxidation promotes hepatic malate-aspartate shuttle activity and glycolysis.

Hepatic metabolic syndrome is associated with inflammation, as inflammation stimulates the reprogramming of nutrient metabolism and hepatic mitochondria-generated acetyl-CoA, but how acetyl-CoA affects the reprogramming of nutrient metabolism, especially glucose and acids, in the condition of inflammation is still unclear. Here, we used an acute inflammation model in which pigs were injected with lipopolysaccharide (LPS) and found that hepatic glycolysis and oxidation are both promoted. Acetyl-proteome profiling of LPS-infected pigs showed that inflammatory stress exacerbates the acetylation of mitochondrial proteins. Both mitochondrial glutamate oxaloacetate transaminase 2 (GOT2) and malate dehydrogenase 2 (MDH2) were acetylated, and the malate-aspartate shuttle (MAS) activity was stimulated to maintain glycolysis. With the use of C-carbon tracing in vitro, acetyl-CoA was found to be mainly supplied by lipid-derived oxidation rather than glucose-derived pyruvate oxidative decarboxylation, while glucose was mainly used for lactate production in response to inflammatory stress. The results of the mitochondrial experiment showed that acetyl-CoA directly increases MDH2 and, in turn, the GOT2 acetylation level affects MAS activity. Treatment with palmitate in primary hepatocytes from LPS-injected pigs increased the hepatic production of acetyl-CoA, pyruvate, and lactate; MAS activity; and hepatic MDH2 and GOT2 hyperacetylation, while the deficiency of long-chain acetyl-CoA dehydrogenase resulted in the stabilization of these parameters. These observations suggest that acetyl-CoA produced by oxidation promotes MAS activity and glycolysis via nonenzymatic acetylation during the inflammatory stress response.

Keyword: fatty liver

plus Synergistically Ameliorates Nonalcoholic Steatohepatitis in HepG2 Cells.

The combination of and radix is frequently prescribed for diseases in TKM. However, the synergic effects of the two herbs on nonalcoholic steatohepatitis (NASH) have not yet been studied. Therefore, we investigated the anti-NASH effects of the water extract of (AI), radix (CL), and combination of the two herbs (ACE). Hepatic steatosis and NASH were induced in HepG2 cells by treatment with (PA, for 6\u2009h) with/without pretreatment of ACE (25 or 50\u2009g/mL), AI (50 or 100\u2009g/mL), CL (50 or 100\u2009g/mL), curcumin (5\u2009g/mL), or scopoletin (5\u2009g/mL). The PA treatment (200\u2009M) drastically altered intracellular triglyceride levels, total cholesterol, and expression levels of genes related to lipid metabolism (CD36, SREBP1c, PPAR-, and PPAR-), whereas pretreatment with ACE significantly attenuated these alterations. ACE also protected HepG2 cells from PA- (300\u2009M-) induced endoplasmic reticulum (ER) stress and apoptosis and attenuated the related key molecules including GRP78, eIF2, and CHOP, respectively. In conclusion, we found synergic effects of and on NASH, supporting the clinical potential for disorders. In addition, modulation of ER stress-relative molecules would be involved in its underlying mechanism.

Keyword: fatty liver

Serum C16:1n7/C16:0 ratio as a diagnostic marker for non-alcoholic steatohepatitis.

Accurate diagnosis of non-alcoholic steatohepatitis (NASH) from non-alcoholic disease (NAFLD) is clinically important. Therefore, there is a need for easier ways of diagnosing NASH. In this study, we investigated the serum composition and evaluated the possibility of using the serum composition as a diagnostic marker of NASH.The subjects were 78 NAFLD patients (non-alcoholic [NAFL]: 30, NASH: 48) and 24 healthy individuals. acids extracted from the tissue and serum were identified and quantified by gas chromatography. In addition, we evaluated the relationship between serum and tissue composition, patient background, and histology. The diagnostic performance of NASH was evaluated by calculating the area under the receiver operating characteristic (AUROC).The results of the analysis showed the C16:1n7/C16:0 ratio to have the strongest correlation between serum and tissue (r\xa0=\xa00.865, P\xa0<\xa00.0001). The serum C16:1n7/C16:0 ratio in the NASH group was higher compared with that in the NAFL group (P\xa0=\xa00.0007). Evaluation of the association of the serum C16:1n7/C16:0 ratio with histology revealed significant correlation with lobular inflammation score, ballooning score, and fibrosis score. The AUROC for predicting NASH in all NAFLD patients was 0.7097. The AUROC was nearly equivalent even when the study subjects were restricted to patients with a fibrosis score\xa0≤\xa02 only (AUROC 0.6917).Measuring the serum C16:1n7/C16:0 ratio may be an effective non-invasive method for diagnosing NASH, particularly in its early stages.© 2019 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.

Keyword: fatty liver

Exogenous Hydrogen Sulfide Alleviates-Induced Intracellular Inflammation in HepG2 Cells.

acids induced hepatic inflammation plays an important role in nonalcoholic disease (NAFLD) pathogenesis. Hydrogen sulfide (HS), an endogenous gasotransmitter, has been established to possess potent anti-inflammation in various human organs. However, the anti-inflammation property of HS in the is still needed to further elucidate. Hence, this study aimed to investigate whether exogenous HS can protect hepatocytes against inflammation induced by (PA). HepG2 hepatocytes were exposed to PA for 24\u2009h to induce free acids-induced inflammation. The cells were pretreated with NaHS (a donor of H2S) before exposure to PA. Cell viability, inflammatory cytokines (TNF-α, IL-6 and IL-1β), NLRP3 inflammasome and NF-κB were measured by a combination of MTT assay, ELISA, Western blot and Immunofluorescence. Here, we found that exogenous HS dose-dependently inhibited the expression of pro-inflammatory cytokines, NLRP3 inflammasome and activation of NF-κB signaling in PA-induced HepG2 cells. Thus, HS might be a candidate therapeutic agent against NAFLD.© Georg Thieme Verlag KG Stuttgart · New York.

Keyword: fatty liver

[Inhibition of chitin oligosaccharide on dyslipidemia and the potential molecular mechanism exploration].

The inhibitory effect of NACOS on dyslipidemia and potential molecular mechanisms by in vitro and in vivo experiments were investigated. For in vitro study, four experimental groups were designed by using HepG2 cells, including the control group, (PA) treatment alone group, NACOS treatment alone group and NACOS + PA treatment group. For in vivo study, male C57BL/6 mice were divided into four groups (n=5) at random including the normal control group (NCD), high fat diet (HFD) group, NACOS treatment alone group, NACOS+HFD group, which were treated for 20 weeks. The used methods in this study were as follows: the observation of lipid droplet deposition in HepG2 cells by oil red O staining, the detection of mRNA levels of lipid metabolism-related regulators and inflammatory cytokine by RT-PCR method, the monitoring of MAPKs and PI3K/Akt pathway activation by Western blotting method. The in vitro study shows that, NACOS had no toxicity on the viability of HepG2 cells at 25-100 μg/mL and significantly reduced the deposition of lipid droplet. Also, based on both in vitro and in vivo investigation, NACOS evidently down-regulated the expression of lipid metabolism-related regulators (PGC1α, Cox5b, Mcad) and inflammatory cytokine (IL-1β) at mRNA level (P<0.05 or 0.01), and suppressed the activation of p38, ERK1/2 and Akt in HepG2 cells and lever tissues from HFD-fed mice (P<0.05 or 0.01). Based on the above, NACOS may inhibit the oxidation of mitochondrial and the lipid biosynthesis, block the inflammatory responses and prevent the HepG2 cells and C57BL/6 mice from lipidemia.

Keyword: fatty liver

Human relaxin-2 attenuates hepatic steatosis and fibrosis in mice with non-alcoholic disease.

Human relaxin-2 reduces hepatic fibrosis in mice. However, the effects of relaxin-2 on hepatic steatosis and fibrosis in animals with non-alcoholic disease (NAFLD) remain to be elucidated. C57BL/6 mice fed a high-fat diet (HFD) or methionine-choline-deficient (MCD) diet were randomly assigned to receive recombinant human relaxin-2 (25 or 75\u2009μg/kg/day) or vehicle for 4 weeks. In HFD-fed mice, relaxin-2 decreased systemic insulin resistance and reduced body weight, epididymal fat mass and serum leptin and insulin concentrations. In livers of HFD-fed mice, relaxin-2 attenuated steatosis and increased phosphorylation of insulin receptor substrate-1, Akt and endothelial nitric oxide synthase (eNOS), and activated genes that regulate oxidation and suppressed acetyl-CoA carboxylase. Relaxin-2 had no direct anti-steatotic effect on primary mouse hepatocytes, but S-nitroso-N-acetylpenicillamine attenuated -induced steatosis and activated genes regulating oxidation in hepatocytes. In mice fed an MCD diet, relaxin-2 attenuated steatosis, inflammation and fibrosis. Relaxin-2 increased eNOS and Akt phosphorylation and transcript levels of cytochrome P450-4a10 and decreased acetyl-CoA carboxylase in MCD-fed mouse livers. Moreover, expression levels of Kupffer cell activation, hepatic stellate cell activation and hepatocyte apoptosis were decreased in MCD diet-fed mice receiving relaxin-2. In conclusion, relaxin-2 reduces hepatic steatosis by activating intrahepatic eNOS in HFD-fed mice and further attenuates fibrosis in MCD diet-fed mice. Therefore, human relaxin-2 is a potential therapeutic treatment for NAFLD.

Keyword: fatty liver

Increased serum concentration of ceramides in obese children with nonalcoholic disease.

Hepatic lipid accumulation is closely related to the development of insulin resistance, which is regarded as one of the most significant risk factors of nonalcoholic disease (NAFLD). Although the exact molecular pathway leading to impaired insulin signaling has not been definitively established, ceramides are suspected mediators of lipid induced hepatic insulin resistance. Therefore, the aim of the study was to evaluate the serum ceramides concentration in obese children with NAFLD.The prospective study included 80 obese children (aged 7-17\xa0years, median 12\xa0years) admitted to our Department to diagnose initially suspected disease. Patients with viral hepatitis (HCV, HBV, CMV), autoimmune (AIH), toxic and metabolic (Wilson\'s disease, alfa-1-antitrypsin deficiency) diseases and celiac disease were excluded. NAFLD was diagnosed based on pediatric diagnostic criteria in obese children with steatosis in ultrasound (US) as well as elevated alanine transaminase (ALT) serum activity after exclusion of other major diseases listed before. Ultrasonography was used as a screening method and for qualitative assessment of the steatosis degree (graded according to Saverymuttu scale). Advanced steatosis was defined as a score\u2009>\u20091. The total intrahepatic lipid content (TILC) was assessed by magnetic resonance proton spectroscopy (HMRS) which is the most accurate technique for assessment of ectopic fat accumulation. Fasting serum concentration of ceramides was measured in 62 children.NAFLD was diagnosed in 31 children. Significant, positive correlation was found between total serum concentration of ceramides and insulin (r\xa0=\u20090.3, p\xa0=\u20090.02) and HOMA-IR (r\xa0=\u20090.28, p\xa0=\u20090.03). Total ceramide concentration as well as specific -ceramides (FA-ceramides) concentrations, namely: myristic, , palmitoleic, stearic, oleic, behenic and lignoceric were significantly higher (p\xa0=\u20090.004, p\xa0=\u20090.003, p\xa0=\u20090.007, p\xa0<\u20090.001, p\xa0=\u20090.035, p\xa0=\u20090.008, p\xa0=\u20090.003, p\xa0=\u20090.006, respectively) in children with NAFLD compared to controls (n\xa0=\u200914). Moreover, children with NAFLD had significantly higher activity of ALT (p\xa0<\u20090.001) and GGT (p\xa0<\u20090.001), HOMA-IR (p\xa0=\u20090.04), BMI (p\xa0=\u20090.046), waist circumference (p\xa0=\u20090.01) steatosis grade in ultrasound (p\xa0<\u20090.001) and TILC in HMRS (p\xa0<\u20090.001) compared to children without NAFLD. We did not find significant differences in total and FA-ceramide species concentrations between children with mild (grade 1) and advanced steatosis in ultrasonography (grade 2-3).Elevated ceramide concentrations in obese patients together with their significant correlation with insulin resistance parameters suggest their association with molecular pathways involved in insulin signaling impairment known to be strongly linked to pathogenesis of non-alcoholic disease.

Keyword: fatty liver

Protein translation associated to PERK arm is a new target for regulation of metainflammation: A connection with hepatocyte cholesterol.

Endoplasmic reticulum stress is a cellular phenomenon that has been associated with metabolic disorders, contributing to the development of obesity, disease, and dyslipidemias. Under metabolic overload conditions, in cells with a high protein-secretory activity, such as hepatocytes and Langerhans β cells, the unfolded protein response (UPR) is critical in to maintain protein homeostasis (proteostasis). UPR integrated by a tripartite signaling system, through activating transcription factor 6, protein kinase R-like endoplasmic reticulum kinase (PERK), and inositol-requiring enzyme 1, regulates gene transcription and translation to resolve stress and conserve proteostasis. In the current study, we demonstrated in hepatocytes under metabolic overload by saturated and stearic acids, through activation of PERK signaling and CCAAT-enhancer-binding protein homologous protein (CHOP) transcription factor, an association with the expression of cyclooxygenase 2. More important, isolated exosomes from supernatants of macrophages exposed to lipopolysaccharides can also induce a metainflammation phenomenon, and when\xa0treated on hepatocytes, induced a rearrangement in cholesterol metabolism through sterol regulatory element-binding protein 2 (SREBP2), low-density lipoprotein receptor (LDLR), apolipoprotein A-I, and ABCA1. Moreover, we demonstrate the cellular effect of terpene-derived molecules, such as cryptotanshinone, isolated of plant Salvia brandegeei, regulating metainflammatory conditions through PERK pathway in both hepatocytes and β cells. Our data suggest the presence of a modulatory mechanism on specific protein translation process. This effect could be mediated by eukaryotic initiation factor-4A, evaluating salubrinal as a control molecule. Likewise, the protective mechanisms of unsaturated acids, such as oleic and palmitoleic were confirmed. Therefore, modulation of metainflammation suggests a new target through PERK signaling in cells with a high secretory activity, and possibly the regulation of cholesterol in hepatocytes is promoted via exosomes.© 2018 Wiley Periodicals, Inc.

Keyword: fatty liver

Melatonin improves insulin resistance and hepatic steatosis through attenuation of alpha-2-HS-glycoprotein.

Melatonin plays an important role in regulating circadian rhythms. It also acts as a potent antioxidant and regulates glucose and lipid metabolism, although the exact action mechanism is not clear. The α2-HS-glycoprotein gene (AHSG) and its protein, fetuin-A (FETUA), are one of the hepatokines and are known to be associated with insulin resistance and type 2 diabetes. The aim of this study was to determine whether melatonin improves hepatic insulin resistance and hepatic steatosis in a FETUA-dependent manner. In HepG2 cells treated with 300\xa0μmol/L of , phosphorylated AKT expression decreased, and FETUA expression increased, but this effect was inhibited by treatment with 10\xa0μmol/L of melatonin. However, melatonin did not improve insulin resistance in FETUA-overexpressing cells, indicating that improvement in insulin resistance by melatonin was dependent on downregulation of FETUA. Moreover, melatonin decreased -induced ER stress markers, CHOP, Bip, ATF-6, XBP-1, ATF-4, and PERK. In addition, in the high-fat diet (HFD) mice, oral treatment with 100\xa0mg/kg/day melatonin for 10\xa0weeks reduced body weight gain to one-third of that of the HFD group and hepatic steatosis. Insulin sensitivity and glucose intolerance improved with the upregulation of muscle p-AKT protein expression. FETUA expression and ER stress markers in the and serum of HFD mice were decreased by melatonin treatment. In conclusion, melatonin can improve hepatic insulin resistance and hepatic steatosis through reduction in ER stress and the resultant AHSG expression.© 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keyword: fatty liver

The BACE1 product sAPPβ induces ER stress and inflammation and impairs insulin signaling.

β-secretase/β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a key enzyme involved in Alzheimer\'s disease that has recently been implicated in insulin-independent glucose uptake in myotubes. However, it is presently unknown whether BACE1 and the product of its activity, soluble APPβ (sAPPβ), contribute to lipid-induced inflammation and insulin resistance in skeletal muscle cells.Studies were conducted in mouse C2C12 myotubes, skeletal muscle from Bace1mice and mice treated with sAPPβ and adipose tissue and plasma from obese and type 2 diabetic patients.We show that BACE1 inhibition or knockdown attenuates palmitate-induced endoplasmic reticulum (ER) stress, inflammation, and insulin resistance and prevents the reduction in Peroxisome Proliferator-Activated Receptor γ Co-activator 1α (PGC-1α) and oxidation caused by palmitate in myotubes. The effects of palmitate on ER stress, inflammation, insulin resistance, PGC-1α down-regulation, and oxidation were mimicked by soluble APPβ in vitro. BACE1 expression was increased in subcutaneous adipose tissue of obese and type 2 diabetic patients and this was accompanied by a decrease in PGC-1α mRNA levels and by an increase in sAPPβ plasma levels of obese type 2 diabetic patients compared to obese non-diabetic subjects. Acute sAPPβ administration to mice reduced PGC-1α levels and increased inflammation in skeletal muscle and decreased insulin sensitivity.Collectively, these findings indicate that the BACE1 product sAPPβ is a key determinant in ER stress, inflammation and insulin resistance in skeletal muscle and gluconeogenesis in .Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Chronic intake of moderate fat-enriched diet induces and low-grade inflammation without obesity in rabbits.

Non-Alcoholic Disease (NAFLD) is the cause of chronic disease. Even though NAFLD is strongly associated with obesity and metabolic syndrome, there is a proportion of patients who develop this condition in the absence of obesity and the underlying mechanisms are poorly understood. We investigated early events in the pathogenesis of non-obese NAFLD, analyzing the impact of the chronic intake of a moderate fat-enriched diet on hepatic lipid accumulation and their relationship with inflammation. Rabbits fed with a moderate -- Enriched Diet 3% (FAED), were evaluated for body weight, biochemical parameters, and function. samples were analyzed by histology and RT-qPCR to measure lipid accumulation, the expression of inflammation-related genes IL-1β, IL-6, IL-10, IL-13, IL-18, COX-2, TNF-α, and TLR-4. Chronic consumption by 6-months of FAED did not generate metabolic changes, but it induced . We also observed the development of low-grade inflammation characterized by the up regulation of TNF-α, IL-13 and IL-18. The consumption by 12-months of FAED caused the overexpression of IL-6, IL-10, IL-13, COX-2, and TLR-4. We show that hepatic steatosis is an early consequence of fat-enriched diets, and that it is accompanied by an immune response that exerts protective effects that prevent the development of metabolic disorders, such as overweight/obesity and metabolic syndrome. However, the excessive intake of acids renders these mechanisms less efficient for delaying the start of metabolic alterations. Rabbits fed with FAED can be used as a model of NAFLD in non-obese and obese groups, especially at early stages of the disease.Copyright © 2019. Published by Elsevier B.V.

Keyword: fatty liver

Stable Isotope-Labeled Lipidomics to Unravel the Heterogeneous Development Lipotoxicity.

Non-alcoholic disease (NAFLD) as a global health problem has clinical manifestations ranging from simple non-alcoholic (NAFL) to non-alcoholic steatohepatitis (NASH), cirrhosis, and cancer. The role of different types of acids in driving the early progression of NAFL to NASH is not understood. Lipid overload causing lipotoxicity and inflammation has been considered as an essential pathogenic factor. To correlate the lipid profiles with cellular lipotoxicity, we utilized (C16:0)- and especially unprecedented palmitoleic (C16:1)-induced lipid overload HepG2 cell models coupled with lipidomic technology involving labeling with stable isotopes. C16:0 induced inflammation and cell death, whereas C16:1 induced significant lipid droplet accumulation. Moreover, inhibition of de novo sphingolipid synthesis by myriocin (Myr) aggravated C16:0 induced lipoapoptosis. Lipid profiles are different in C16:0 and C16:1-treated cells. Stable isotope-labeled lipidomics elucidates the roles of specific acids that affect lipid metabolism and cause lipotoxicity or lipid droplet formation. It indicates that not only saturation or monounsaturation of acids plays a role in hepatic lipotoxicity but also Myr inhibition exasperates lipoapoptosis through ceramide in-direct pathway. Using the techniques presented in this study, we can potentially investigate the mechanism of lipid metabolism and the heterogeneous development of NAFLD.

Keyword: fatty liver

Interesterified Fats Induce Deleterious Effects on Adipose Tissue and in LDLr-KO Mice.

Interesterified fats are being widely used by the food industry in an attempt to replace trans acids. The effect of interesterified fats containing or stearic acids on lipid metabolism and inflammatory signaling pathways in adipose and hepatic tissues was evaluated. Male LDLr-KO mice were fed a high-fat diet containing polyunsaturated (PUFA), (PALM), interesterified (PALM INTER), stearic (STEAR), or stearic interesterified (STEAR INTER) fats for 16 weeks. The expression of genes and protein levels involved in lipid metabolism and inflammatory processes in and white adipose tissue was determined by quantitative RT-PCR and by Western blot, respectively. The infiltration of inflammatory cells in hepatic and adipose tissues was determined by eosin and hematoxylin, while collagen content was determined by Sirius Red staining. Both interesterified fats increased collagen content and JNK phosphorylation. Additionally, the STEAR INTER group developed nonalcoholic steatohepatitis (NASH) associated with higher neutrophil infiltration. PALM INTER induced adipose tissue expansion and enlargement of adipocytes. Furthermore, PALM INTER triggered increased IKK phosphorylation and TNFα protein content, conditions associated with the upstream activation of the NFkB signaling pathway. STEAR INTER induced NASH, while PALM INTER triggered hepatic fibrosis and adipocyte hypertrophy with inflammatory response in LDLr-KO mice.

Keyword: fatty liver

Dynamic alterations in the gut microbiota and metabolome during the development of methionine-choline-deficient diet-induced nonalcoholic steatohepatitis.

To investigate changes in gut microbiota and metabolism during nonalcoholic steatohepatitis (NASH) development in mice fed a methionine-choline-deficient (MCD) diet.Twenty-four male C57BL/6J mice were equally divided into four groups and fed a methionine-choline-sufficient diet for 2 wk (Control 2w group, = 6) or 4 wk (Control 4w group, = 6) or the MCD diet for 2 wk (MCD 2w group, = 6) or 4 wk (MCD 4w group, = 6). injury, fibrosis, and intestinal barrier function were evaluated after 2 and 4 wk of feeding. The fecal microbiome and metabolome were studied using 16s rRNA deep sequencing and gas chromatography-mass spectrometry.The mice fed the MCD diet presented with simple hepatic steatosis and slight intestinal barrier deterioration after 2 wk. After 4 wk of feeding with the MCD diet, however, the mice developed prominent NASH with fibrosis, and the intestinal barrier was more impaired. Compared with the control diet, the MCD diet induced gradual gut microbiota dysbiosis, as evidenced by a marked decrease in the abundance of and the () group ( < 0.001 and < 0.05, respectively) and a significant increase in Ruminococcaceae UCG 014 abundance ( < 0.05) after 2 wk. At 4 wk, the MCD diet significantly reduced the promising probiotic levels and markedly promoted abundance ( < 0.05, and < 0.01, respectively). The fecal metabolomic profile was also substantially altered by the MCD diet: At 2 wk, arachidic , hexadecane, , and tetracosane were selected as potential biomarkers that were significantly different in the corresponding control group, and at 4 wk, cholic , cholesterol, arachidic , tetracosane, and stearic were selected.The MCD diet induced persistent alterations in the gut microbiota and metabolome.

Keyword: fatty liver

Hepatocyte growth factor alleviates hepatic insulin resistance and lipid accumulation in high-fat diet-fed mice.

Type 2 diabetes mellitus is frequently accompanied by disease. Lipid accumulation within the is considered as one of the risk factors for insulin resistance. Hepatocyte growth factor (HGF) is used to treat dysfunction; however, the effect and mechanism of HGF on hepatic lipid metabolism are still not fully understood.Male C57BL/6 mice were induced with a high-fat diet for 12 weeks, followed by a 4-week treatment of HGF or vehicle saline. The levels of fasting blood glucose, fasting insulin and homeostatic model assessment of insulin resistance were calculated for insulin sensitivity. Biochemical plasma parameters were also measured to assess the effect of HGF on lipid accumulation. Additionally, genes in the lipid metabolism pathway were evaluated in -treated HepG2 cells and high-fat diet mice.HGF treatment significantly decreased the levels of fasting blood glucose, hepatic triglyceride and cholesterol contents. Additionally, HGF-regulated expression levels of sterol regulatory element-binding protein-1c/ synthase, peroxidase proliferator-activated receptor-α, and upstream nuclear receptors, such as farnesoid X receptor and small heterodimer partner. Furthermore, c-Met inhibitor could partially reverse the effects of HGF.HGF treatment can ameliorate hepatic insulin resistance and steatosis through regulation of lipid metabolism. These effects might occur through farnesoid X receptor-small heterodimer partner axis-dependent transcriptional activity.© 2018 The Authors. Journal of Diabetes Investigation published by Asian Association for the Study of Diabetes (AASD) and John Wiley & Sons Australia, Ltd.

Keyword: fatty liver

C-X-C Motif Chemokine 10 Impairs Autophagy and Autolysosome Formation in Non-alcoholic Steatohepatitis.

C-X-C motif chemokine 10 (CXCL10) is a crucial pro-inflammatory factor in chronic hepatitis. Autophagy dysregulation is known to contribute to hepatic inflammatory injury. Hence, we investigated the regulatory effect of CXCL10 on the autophagosome-lysosome system during non-alcoholic disease (NAFLD) development. The effect of CXCL10 ablation by neutralizing monoclonal antibody (mAb) or genetic knockout on autophagic flux was evaluated in cultured hepatocytes and animal models of NAFLD. Results demonstrated that CXCL10 ablation protected against hepatocyte injury and steatohepatitis development in mice. Autophagic flux impairment was rectified by CXCL10 inhibition using anti-CXCL10 mAb in AML-12 and HepG2 cell lines and primary hepatocytes as evidenced by the attenuated accumulation of p62/SQSTM1 and LC3-II proteins and increased autophagic protein degradation. Impaired autophagic flux was significantly restored by CXCL10 knockout or anti-CXCL10 mAb in mice. Bafilomycin A1, an inhibitor of autolysosome formation, abolished the rectifying effect of anti-CXCL10 mAb or CXCL10 knockdown in AML-12 and primary hepatocytes, indicating CXCL10 impaired late-stage autophagy in NAFLD. Anti-CXCL10 mAb treatment also increased the fusion of LC3-positive autophagosomes with lysosomes in HepG2 cells challenged with , suggesting that CXCL10 ablation restored autolysosome formation. Consistently, the number of autolysosomes was significantly increased by CXCL10 knockout in mice as shown by electron microscopy. In conclusion, upregulated CXCL10 in steatohepatitis impairs autophagic flux by reducing autolysosome formation, thereby inhibiting autophagic protein degradation and the accumulation of ubiquitinated proteins, leading to the development of steatohepatitis.

Keyword: fatty liver

Lipotoxicity and the gut- axis in NASH pathogenesis.

The pathogenesis of non-alcoholic disease, particularly the mechanisms whereby a minority of patients develop a more severe phenotype characterised by hepatocellular damage, inflammation, and fibrosis is still incompletely understood. Herein, we discuss two pivotal aspects of the pathogenesis of NASH. We first analyse the initial mechanisms responsible for hepatocellular damage and inflammation, which derive from the toxic effects of excess lipids. Accumulating data indicate that the total amount of triglycerides stored in hepatocytes is not the major determinant of lipotoxicity, and that specific lipid classes act as damaging agents on cells. In particular, the role of free acids such as , cholesterol, lysophosphatidylcholine and ceramides has recently emerged. These lipotoxic agents affect the cell behaviour via multiple mechanisms, including activation of signalling cascades and death receptors, endoplasmic reticulum stress, modification of mitochondrial function, and oxidative stress. In the second part of this review, the cellular and molecular players involved in the cross-talk between the gut and the are considered. These include modifications to the microbiota, which provide signals through the intestine and bacterial products, as well as hormones produced in the bowel that affect metabolism at different levels including the . Finally, the activation of nuclear receptors by bile acids is analysed.Copyright © 2017. Published by Elsevier B.V.

Keyword: fatty liver

Attenuated lipotoxicity and apoptosis is linked to exogenous and endogenous augmenter of regeneration by different pathways.

Nonalcoholic disease (NAFLD) covers a spectrum from simple steatosis to nonalcoholic steatohepatitis (NASH) and cirrhosis. Free acids (FFA) induce steatosis and lipo-toxicity and correlate with severity of NAFLD. In this study we aimed to investigate the role of exogenous and endogenous ALR (augmenter of regeneration) for FFA induced ER (endoplasmatic reticulum) -stress and lipoapoptosis. Primary human hepatocytes or hepatoma cells either treated with recombinant human ALR (rhALR, 15kDa) or expressing short form ALR (sfALR, 15kDa) were incubated with (PA) and analyzed for lipo-toxicity, -apoptosis, activation of ER-stress response pathways, triacylglycerides (TAG), mRNA and protein expression of lipid metabolizing genes. Both, exogenous rhALR and cytosolic sfALR reduced PA induced caspase 3 activity and Bax protein expression and therefore lipotoxicity. Endogenous sfALR but not rhALR treatment lowered TAG levels, diminished activation of ER-stress mediators C-Jun N-terminal kinase (JNK), X-box binding protein-1 (XBP1) and proapoptotic transcription factor C/EBP-homologous protein (CHOP), and reduced death receptor 5 protein expression. Cellular ALR exerts its lipid lowering and anti-apoptotic actions by enhancing FABP1, which binds toxic FFA, increasing mitochondrial β-oxidation by elevating the mitochondrial FFA transporter CPT1α, and decreasing ELOVL6, which delivers toxic FFA metabolites. We found reduced hepatic mRNA levels of ALR in a high fat diet mouse model, and of ALR and FOXA2, a transcription factor inducing ALR expression, in human steatotic as well as NASH samples, which may explain increased lipid deposition and reduced β-oxidation in NASH patients. Present study shows that exogenous and endogenous ALR reduce PA induced lipoapoptosis. Furthermore, cytosolic sfALR changes mRNA and protein expression of genes regulating lipid metabolism, reduces ER-stress finally impeding progression of NASH.

Keyword: fatty liver

Anticandidal activity of the extract and compounds isolated from Rottb.

The phytochemical screening of showed that carbohydrates and/or glycosides, flavonoids, tannins, sterols and/or triterpenes, and proteins and/or amino acids are present. The profile comprised major; , oleic, heptadecanoic, linoleic and minor; arachidonic, lignoceric, stearic, and myristic . Two compounds; namely, α-amyrin and β-sitosterol were isolated by the fractionation of unsaponifiable matter. The acute toxicity study showed that the reported after oral administration of the alcohol extract ( showed that the plant was highly safe as the LD was more than 4000\u202fmg/kg. These results were well supported by the sub-chronic toxicity, as the administrated to rats for 15 consecutive days at dose 1000\u202fmg/kg showed no alteration in the and kidney functions. Moreover, the extract of the plant exhibited anti-candidal activity against different species. The most potent activity, (23.1\u202f±\u202f2.1, 0.98\u202fµg/ml) and (22.3\u202f±\u202f0.53, 0.98\u202fµg/ml), was obtained by the chloroform and total extract, respectively against .

Keyword: fatty liver

ACSL5 genotype influence on metabolism: a cellular, tissue, and whole-body study.

Acyl-CoA Synthetase Long Chain 5 (ACSL5) gene\'s rs2419621 T/C polymorphism was associated with ACSL5 mRNA expression and response to lifestyle interventions. However, the mechanistic understanding of the increased response in T allele carriers is lacking. Study objectives were to investigate the effect of rs2419621 genotype and ACSL5 human protein isoforms on oxidation and respiration.Human ACSL5 overexpression in C2C12 mouse myoblasts was conducted to measure C oxidation and protein isoform localization in vitro. C oxidation studies and Western blot analysis of ACSL5 proteins were carried out in rectus abdominis primary myotubes from 5 rs2419621 T allele carriers and 4 non-carriers. In addition, mitochondrial high-resolution respirometry was conducted on vastus lateralis muscle biopsies from 4 rs2419621 T allele carriers and 4 non-carriers. Multiple linear regression analysis was conducted to test the association between rs2419621 genotype and respiratory quotient related pre- and post-lifestyle intervention measurements in postmenopausal women with overweight or obesity.In comparison to rs2419621 non-carriers, T allele carriers displayed higher levels of i) 683aa ACSL5 isoform, localized mainly in the mitochondria, playing a greater role in oxidation in comparison to the 739aa protein isoform ii) in vitro CO production in rectus abdominis primary myotubes iii) in vivo oxidation and lower carbohydrate oxidation post-intervention iv) ex vivo complex I and II tissue respiration in vastus lateralis muscle.These results support the conclusion that rs2419621 T allele carriers, are more responsive to lifestyle interventions partly due to an increase in the short ACSL5 protein isoform, increasing cellular, tissue and whole-body utilization. With the increasing effort to develop personalized medicine to combat obesity, our findings provide additional insight into genotypes that can significantly affect whole body metabolism and response to lifestyle interventions.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Loss of toll-like receptor 3 aggravates hepatic inflammation but ameliorates steatosis in mice.

The importance of toll-like receptor (TLR) 4 in the pathogenesis of steatohepatitis has been well documented; however, little is known about the role of TLR3. In this study, we determined whether the depletion of TLR3 modulated hepatic injury in mice and further aimed to provide mechanistic insights into the TLR3-mediated modulation of diet-induced hepatic inflammation and fat accumulation. Hepatic steatosis and inflammatory response were induced by feeding wild-type (WT) or TLR3 knockout mice a high-fat diet for 8 weeks. Primary resident cells, including hepatocytes, Kupffer cells, and hepatic stellate cells (HSCs), were treated with . TLR3 knockout mice fed a high-fat diet showed severe hepatic inflammation accompanied by nuclear factor-κB and IRF3 activation, which is mainly induced by the activation of Kupffer cells. Decreased TLR4 expression was restored in hepatic mononuclear cells and Kupffer cells in TLR3 knockout mice compared to that in the WT. Moreover, hepatic steatosis was decreased in TLR3 knockout mice. Hepatocytes from TLR3 knockout mice exhibited reduced expression of cannabinoid receptors. HSCs from TLR3 knockout mice showed decreased expression of the enzymes involved in endocannabinoid synthesis. In conclusion, this study suggests that the selective modulation of TLR3 could be a novel therapeutic target for the treatment of hepatic inflammation and steatosis.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Tannic , a novel histone acetyltransferase inhibitor, prevents non-alcoholic disease both in\xa0vivo and in\xa0vitro model.

We examined the potential of tannic (TA) as a novel histone acetyltransferase inhibitor (HATi) and demonstrated that TA prevents non-alcoholic disease (NAFLD) by inhibiting HAT activity.The anti-HAT activity of TA was examined using HAT activity assays. An in\xa0vitro NAFLD model was generated by treating HepG2 cells with oleic and acids. Male C57BL/6J mice were fed a control diet (CD) or Western diet (WD) with or without supplementation with either 1% or 3% TA (w/w) for 12 weeks. Finally, the possibility of interacting p300 and TA was simulated.TA suppressed HAT activity both in\xa0vitro and in\xa0vivo. Interestingly, TA abrogated occupancy of p300 on the sterol regulatory element in the synthase and ATP-citrate lyase promoters, eventually inducing hypoacetylation of H3K9 and H3K36. Furthermore, TA decreased acetylation at lysine residues 9 and 36 of histone H3 protein and that of total proteins. Consequently, TA decreased the mRNA expression of lipogenesis-related genes and attenuated lipid accumulation in\xa0vivo. We observed that NAFLD features, including body weight, mass, fat mass, and lipid profile in serum, were improved by TA supplementation in\xa0vivo. Finally, we demonstrated the possibility that TA directly binds to p300 through docking simulation between ligand and protein.Our findings demonstrate that TA, a novel HATi, has potential application for the prevention of NAFLD.Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.

Keyword: fatty liver

Autophagy activation by Jiang Zhi Granule protects against metabolic stress-induced hepatocyte injury.

To elucidate the potential role of autophagy and the protective effects of Jiang Zhi Granule (JZG) in metabolic stress-induced hepatocyte injury.An and approach was used in this study. HepG2 cells were incubated in culture medium containing palmitate (PA; 0, 0.1, 0.2, 0.3, 0.4 or 0.5 mmol/L) and treated with or without JZG (100 μg/mL) for 24 h or 48 h, and the progression of autophagy was visualized by stable fluorescence-expressing cell lines LC3 and p62. Western blot analyses were performed to examine the expression of LC3-II/LC3-I, p62, mTOR and PI3K, while mitochondrial integrity and oxidative stress were observed by fluorescence staining of JC-1 and reactive oxygen species. C57BL/6 mice were divided into three groups: control group ( = 10), high fat (HF) group ( = 13) and JZG group ( = 13); and, histological staining was carried out to detect inflammation and lipid content in the .The cell trauma induced by PA was aggravated in a dose- and time-dependent manner, and hepatic function was improved by JZG. PA had dual effects on autophagy by activating autophagy induction and blocking autophagic flux. The PI3K-AKT-mTOR signaling pathway and the fusion of isolated hepatic autophagosomes and lysosomes were critically involved in this process. JZG activated autophagy progression by either induction of autophagosomes or co-localization of autophagosomes and lysosomes as well as degradation of autolysosomes to protect against PA-induced hepatocyte injury, and protected mitochondrial integrity against oxidative stress in PA-induced mitochondrial dysfunction. In addition, JZG ameliorated lipid droplets and inflammation induced by HF diet , leading to improved metabolic disorder and associated injury in a mouse model of non-alcoholic disease (NAFLD).Metabolic stress-induced hepatocyte injury exhibited dual effects on autophagy and JZG activated the entire process, resulting in beneficial effects in NAFLD.

Keyword: fatty liver

Role of 3-Hydroxy -Induced Hepatic Lipotoxicity in Acute of Pregnancy.

Acute of pregnancy (AFLP), a catastrophic illness for both the mother and the unborn offspring, develops in the last trimester of pregnancy with significant maternal and perinatal mortality. AFLP is also recognized as an obstetric and medical emergency. Maternal AFLP is highly associated with a fetal homozygous mutation (1528G>C) in the gene that encodes for mitochondrial long-chain hydroxy acyl-CoA dehydrogenase (LCHAD). The mutation in LCHAD results in the accumulation of 3-hydroxy acids, such as 3-hydroxy myristic , 3-hydroxy and 3-hydroxy dicarboxylic in the placenta, which are then shunted to the maternal circulation leading to the development of acute injury observed in patients with AFLP. In this review, we will discuss the mechanistic role of increased 3-hydroxy in causing lipotoxicity to the and in inducing oxidative stress, mitochondrial dysfunction and hepatocyte lipoapoptosis. Further, we also review the role of 3-hydroxy acids in causing placental damage, pancreatic islet β-cell glucolipotoxicity, brain damage, and retinal epithelial cells lipoapoptosis in patients with LCHAD deficiency.

Keyword: fatty liver

Lycopus lucidus Turcz. ex Benth. Attenuates free -induced steatosis in HepG2 cells and non-alcoholic disease in high-fat diet-induced obese mice.

Non-alcoholic disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance.We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD.An in vitro modelwas established by treatment of HepG2 cells with a 1\u202fmM free (FFA) mixture (oleic /, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200\u202f\xa0mg/kg daily by oral gavage).HepG2 cells were exposed to 1\u202fmM FFA, with or without LLE (250 -\xa01000\u202f\xa0mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the . Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid metabolism.LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice.These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.Copyright © 2018. Published by Elsevier GmbH.

Keyword: fatty liver

High-fat diet promotes experimental by inducing oxidative stress in the colon.

Diets high in animal fats are associated with increased risks of inflammatory bowel disease, but the mechanism remains unclear. In this study, we investigated the effect of high-fat diet (HFD) on the development of experimental in mice. Relative to mice fed low-fat diet (LFD), HFD feeding for 4 wk increased the levels of triglyceride, cholesterol, and free fatty acids in the plasma as well as within the colonic mucosa. In an experimental model induced by 2,4,6-trinitrobenzenesulfonic (TNBS), mice on 4-wk HFD exhibited more severe colonic inflammation and developed more severe compared with the LFD counterparts. HFD feeding resulted in higher production of mucosal pro-inflammatory cytokines, greater activation of the myosin light chain kinase (MLCK) tight junction regulatory pathway, and greater increases in mucosal barrier permeability in mice following TNBS induction. HFD feeding also induced gp91, an NADPH oxidase subunit, and promoted reactive oxygen species (ROS) production in both colonic epithelial cells and lamina propria cells. In HCT116 cell culture, or and TNF-α combination markedly increased ROS production and induced the MLCK pathway, and these effects were markedly diminished in the presence of a ROS scavenger. Taken together, these data suggest that HFD promotes by aggravating mucosal oxidative stress, which rapidly drives mucosal inflammation and increases intestinal mucosal barrier permeability. This study demonstrates high-fat diet feeding promotes in a 2,4,6-trinitrobenzenesulfonic -induced experimental model in mice. The underlying mechanism is that high-fat diet induces oxidative stress in the colonic mucosa, which increases colonic epithelial barrier permeability and drives colonic mucosal inflammation. These observations provide molecular evidence that diets high in saturated fats are detrimental to patients with inflammatory bowel diseases.

Keyword: fatty liver

Involvement of sarco/endoplasmic reticulum calcium ATPase-mediated calcium flux in the protective effect of oleic against lipotoxicity in hepatocytes.

Elevated free acids, particularly saturated ones such as , may play an important role in the lipotoxic mechanism of nonalcoholic disease (NAFLD). Saturated acids induce autophagy dysfunction and endoplasmic reticulum (ER) stress leading to apoptosis in hepatocytes. However, unsaturated acids, such as oleic , are nontoxic and can even prevent saturated -induced toxicity in vitro. Although emerging evidence has suggested that ER calcium flux disruption in hepatocytes is involved in NAFLD pathogenesis, the roles of acids in autophagy and ER calcium flux still remain unclear. We demonstrated that oleic ameliorated -induced autophagy arrest and ER stress in parallel with ER calcium depletion in hepatocytes. Moreover, we found that the effect of oleic against autophagy arrest was reversed by the pharmacological inhibition of sarcoplasmic reticulum Ca-ATPase (SERCA), which influxes calcium to ER. These data suggest that SERCA-mediated ER calcium flux is greatly involved in -induced lipotoxicity in hepatocytes, and the prevention of ER calcium depletion may restore saturated -induced autophagy arrest in hepatocytes.Copyright © 2019. Published by Elsevier Inc.

Keyword: fatty liver

Adiponectin homolog novel osmotin protects obesity/diabetes-induced NAFLD by upregulating AdipoRs/PPARα signaling in ob/ob and db/db transgenic mouse models.

In metabolic disorders, adiponectin and adiponectin receptors (AdipoR1/R2) signaling has a key role in improving nonalcoholic disease (NAFLD) in obesity-associated diabetes.To the best of our knowledge, here, we reported for the first time the underlying mechanistic therapeutic efficacy of the novel osmotin, a homolog of mammalian adiponectin, against NAFLD in leptin-deficient ob/ob and db/db mice.The ob/ob and db/db mice were treated with osmotin at a dose of 5\u202fμg/g three times a week for two weeks. To co-relate the in vivo results we used the human carcinoma HepG2 cells, subjected to knockdown with small siRNAs of AdipoR1/R2 and PPARα genes and treated with osmotin and (P.A.). MTT assay, Western blotting, immunohistofluorescence assays, and plasma biochemical analyses were applied.Osmotin stimulated AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways in ob/ob and db/db mice, and HepG2 cells exposed to P.A. Mechanistically, we confirmed that knockdown of AdipoR1/R2 and PPARα by their respective siRNAs abolished the osmotin activity in HepG2 cells exposed to P.A. Overall, the in vivo and in vitro results suggested that osmotin protected against NAFLD through activation of AdipoR1/R2 and its downstream APPL1/PPAR-α/AMPK/SIRT1 pathways as shown by the reduced body weight, blood glucose level and glycated hemoglobin, improved glucose tolerance, attenuated insulin resistance and hepatic glucogenesis, regulated serum lipid parameters, and increased oxidation and mitochondrial functions.Our findings strongly suggest that novel osmotin might be a potential novel therapeutic tool against obesity/diabetes-induced NAFLD and other metabolic disorders.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Impact of miR-140 Deficiency on Non-Alcoholic Disease.

We have previously shown that loss of miR-140 has a pro-fibrotic effect in the mammary gland. This study aims to investigate whether miR-140 loss and obesity act synergistically to promote non-alcoholic disease (NAFLD), and to identify the underlying mechanisms. tissues were isolated from lean-fat-diet and high-fat-diet fed wild-type and miR-140 knockout mice. Using molecular staining and immunohistochemistry techniques, increased development of NAFLD and fibrotic indicators in miR-140 knockout mice were identified. Utilizing an in vitro model system, miR-140 was demonstrated to target TLR-4, and miR-140 overexpression was shown to be sufficient to inhibit signaling through the TLR-4/NFκB pathway.These findings demonstrate that loss of miR-140 results in increased expression of TLR-4, sensitizing cells to signaling and in increased inflammatory activity through the TLR4/NFκB pathway. This signaling axis promotes NAFLD development in a high-fat diet context and indicates the potential utility of miR-140 rescue as a therapeutic strategy in NAFLD.© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keyword: fatty liver

Hydroxyhydroquinone impairs fat utilization in mice by reducing nitric oxide availability.

Habitual consumption of chlorogenic compounds (CGAs) from coffee increases fat catabolism and reduces body fat; however, the contribution of roasted coffee remains unclear. Hydroxyhydroquinone (HHQ) impairs the vasodilatory and antihypertensive effects of CGAs by reducing nitric oxide (NO) bioavailability. Since HHQ also reduces fat catabolism, we hypothesized that HHQ does so by decreasing NO availability. Therefore, we investigated the effect of HHQ on energy metabolism in KKAy mice. In HHQ-treated mice, fat oxidation was significantly low and dose-dependent, serum and urinary hydrogen peroxide were high, and plasma NO metabolites and S-nitrosylated proteins were low. In HHQ-treated mouse hepatocytes, the palmitate-induced increase in cellular oxygen consumption was negatively affected, and HHQ or L-NAME reduced cellular utilization. In conclusion, HHQ can impair fat utilization by reducing NO availability in mice. Protein S-nitrosylation reduction in cells after HHQ consumption may be associated with impaired oxidation.

Keyword: fatty liver

PPARγ alleviated hepatocyte steatosis through reducing SOCS3 by inhibiting JAK2/STAT3 pathway.

Peroxisome proliferator-activated receptor gamma (PPARγ) participates in the process of insulin resistance (IR), a crucial pathophysiology in non-alcoholic disease (NAFLD). Meanwhile, suppressor of cytokine signaling3 (SOCS3) also regulates IR in NAFLD. Both PPARγ and SOCS3 play a role in NAFLD through regulating IR, while it is unclear whether these two proteins interact to regulate hepatic steatosis. PPARγ, SOCS3 and its associated JAK2/STAT3 pathway were analyzed using Kuppfer cells (KCs) treatment with LPS and BRL-3A cells treatment with , KC-conditioned medium (KCCM), PPARγ agonist rosiglitazone (ROZ) or JAK2 inhibitor AG490 to demonstrate the role of PPARγ and SOCS3 in hepatocytes steatosis. As LPS concentration increasing, phagocytosis activity of KCs decreased; but releasing of TNF-α and IL-6 increased. After treatment with KCCM, mRNA level of SOCS3, JAK2 and STAT3 as well as protein expression of SOCS3, p-JAK2 and p-STAT3 in steatosis BRL-3A cells increased significantly, which were inhibited by AG490 or ROZ treatment. Taken together, these results indicated that KCCM attributed to KCs dysfunction facilitated hepatocyte steatosis through promoting expressing SOCS3; but PPARγ agonist ROZ alleviated steatosis through reducing SOCS3 expression by inhibiting JAK2/STAT3 in hepatocytes.Copyright © 2018 Elsevier Inc. All rights reserved.

Keyword: fatty liver

Bromide alleviates -induced lipid accumulation in mouse primary hepatocytes through the activation of PPARα signals.

Increased plasma free acids (FFAs) and triglyceride (TG) accumulations have been implicated in the pathogenesis of hepatic steatosis. On the other hand, trace elements function as essential cofactors that are involved in various biochemical processes in mammals, including metabolic homeostasis. Notably, clinical and animal studies suggest that the plasma levels of bromide negatively correlate with those of TG, total cholesterol (TC) and high-density lipoprotein-cholesterol (HDL-C). However, the effect of bromide on lipid accumulation and the direct molecular target responsible for its action remains unknown. Oil red O (ORO) and Nile red staining were used to detect the effect of bromide on lipid accumulation in mouse primary hepatocytes (PHs) treated with different doses of sodium bromide (NaBr) in the presence of FFAs (0.4\xa0mM oleate/ 1:1). Spectrophotometric and fluorometric analyses were performed to assess cellular TG concentrations and rates of oxidation (FAO), respectively, in mouse PHs. We found that bromide decreased FFA-induced lipid accumulation and increased FFA-inhibited oxygen consumptions in mouse PHs in a dose-dependent manner via activation of PPARα. Mechanical studies demonstrated that bromide decreased the phosphorylation levels of JNK. More importantly, the PPARα-specific inhibitor GW6471 partially abolished the beneficial effects of bromide on mouse PHs. Bromide alleviates FFA-induced excessive lipid storage and increases rates of FAO through the activation of PPARα/JNK signals in mouse PHs. Therefore, bromide may serve as a novel drug in the treatment of hepatic steatosis.© 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Keyword: fatty liver

miR-1224-5p Enhances Hepatic Lipogenesis by Targeting Adenosine Monophosphate-Activated Protein Kinase α1 in Male Mice.

MicroRNAs are potential therapeutic targets for metabolic diseases. Here, miR-1224-5p was highly expressed in the livers of mice fed a high-fat diet (HFD) and in obese (ob/ob) mice. To examine the potential role of miR-1224-5p, we constructed -specific adenoviral vectors expressing either an miR-1224-5p inhibitor sequence or miR-1224-5p mimic sequences. After tail-vein vector injection, HFD-fed mice were examined for expression of lipogenic genes. We found that miR-1224-5p inhibitors significantly attenuated hepatic lipogenesis and steatosis in HFD-fed mice, whereas miR-1224-5p mimicked promoted lipid accumulation in the of chow-fed C57BL/6 mice. Additional in vitro studies demonstrated that downregulation of miR-1224-5p in HepG2 and primary hepatocytes led to a reduction of cellular triglycerides after treatment with an oleic and mixture. Importantly, this study also identified adenosine monophosphate-activated protein kinase (AMPK)-α1 as a direct target of miR-1224-5p. miR-1224-5p binding to the 3\' untranslated region of AMPKα1 suppressed expression of the AMPKα1 protein and its downstream molecules. Metformin, an activator of AMPK, also inhibited hepatic expression of miR-1224-5p. Together, these findings indicate that miR-1224-5p promotes hepatic lipogenesis by suppressing AMPKα1 expression and suggest that miR-1224-5p inhibitors warrant further investigation as potential therapeutic tools in the treatment of nonalcoholic disease.

Keyword: fatty liver

Pro-inflammatory hepatic macrophages generate ROS through NADPH oxidase 2 via endocytosis of monomeric TLR4-MD2 complex.

Reactive oxygen species (ROS) contribute to the development of non-alcoholic disease. ROS generation by infiltrating macrophages involves multiple mechanisms, including Toll-like receptor 4 (TLR4)-mediated NADPH oxidase (NOX) activation. Here, we show that palmitate-stimulated CD11bF4/80 hepatic infiltrating macrophages, but not CD11bF4/80 Kupffer cells, generate ROS via dynamin-mediated endocytosis of TLR4 and NOX2, independently from MyD88 and TRIF. We demonstrate that differently from LPS-mediated dimerization of the TLR4-MD2 complex, palmitate binds a monomeric TLR4-MD2 complex that triggers endocytosis, ROS generation and increases pro-interleukin-1β expression in macrophages. Palmitate-induced ROS generation in human CD68CD14 macrophages is strongly suppressed by inhibition of dynamin. Furthermore, Nox2-deficient mice are protected against high-fat diet-induced hepatic steatosis and insulin resistance. Therefore, endocytosis of TLR4 and NOX2 into macrophages might be a novel therapeutic target for non-alcoholic disease.

Keyword: fatty liver

Hepatocyte nuclear receptor SHP suppresses inflammation and fibrosis in a mouse model of nonalcoholic steatohepatitis.

Nonalcoholic disease (NAFLD) is a burgeoning health problem worldwide, ranging from nonalcoholic (NAFL, steatosis without hepatocellular injury) to the more aggressive nonalcoholic steatohepatitis (NASH, steatosis with ballooning, inflammation, or fibrosis). Although many studies have greatly contributed to the elucidation of NAFLD pathogenesis, the disease progression from NAFL to NASH remains incompletely understood. Nuclear receptor small heterodimer partner (Nr0b2, ) is a transcriptional regulator critical for the regulation of bile , glucose, and lipid metabolism. Here, we show that SHP levels are decreased in the livers of patients with NASH and in diet-induced mouse NASH. Exposing primary mouse hepatocytes to and lipopolysaccharide , we demonstrated that the suppression of expression in hepatocytes is due to c-Jun N-terminal kinase (JNK) activation, which stimulates c-Jun-mediated transcriptional repression of Interestingly, induction of hepatocyte-specific SHP in steatotic mouse ameliorated NASH progression by attenuating inflammation and fibrosis, but not steatosis. Moreover, a key mechanism linking the anti-inflammatory role of hepatocyte-specific SHP expression to inflammation involved SHP-induced suppression of NF-κB p65-mediated induction of chemokine (C-C motif) ligand 2 (CCL2), which activates macrophage proinflammatory polarization and migration. In summary, our results indicate that a JNK/SHP/NF-κB/CCL2 regulatory network controls communications between hepatocytes and macrophages and contributes to the disease progression from NAFL to NASH. Our findings may benefit the development of new management or prevention strategies for NASH.© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

Keyword: fatty liver

Angiotensin-converting enzyme 2 inhibits endoplasmic reticulum stress-associated pathway to preserve nonalcoholic disease.

Previous works indicated that the stress on the endoplasmic reticulum (ER) affected nonalcoholic disease (NAFLD). However, there is no clear evident on the effect of the regulation of ER stress by angiotensin-converting enzyme 2 (ACE2) on the prevention of NAFLD.HepG2 cells were treated with thapsigargin (Tg) or (PA). We analysed ACE2 expression using Western-blotting analyses. ER stress-related proteins were detected in ACE2 knockout mice and Ad-ACE2-treated db/db mice by immunofluorescence or Western-blotting analyses. In ACE2-overexpression HepG2 cells, the triglyceride (TG), total cholesterol (TC), and glycogen content were detected by assay kits. Meanwhile, the expression of hepatic lipogenic proteins (ACCα, SREBP-1c, FAS, and LXRα), enzymes for gluconeogenesis (PEPCK, G6Pase, and IRS2), and IKKβ/NFκB/IRS1/Akt pathway were analysed by Western-blotting analyses.ACE2 was significantly increased in Tg/PA-induced cultured hepatocytes. Additionally, ACE2 knockout mice displayed elevated levels of ER stress, while Ad-ACE2-treated db/db mice showed reduced ER stress in . Furthermore, activation of ACE2 can ameliorate ER stress, accompanied by decreased TG content, increased intracellular glycogen, and downregulated expression of hepatic lipogenic proteins and enzymes for gluconeogenesis in Tg/PA-induced hepatocytes. As a consequence of anti-ER stress, the activation of ACE2 led to improved glucose and lipid metabolism through the IKKβ/NFκB/IRS1/Akt pathway.This is the first time documented that ACE2 had a notable alleviating role in ER stress-induced hepatic steatosis and glucose metabolism via the IKKβ/NFκB/IRS1/Akt-mediated pathway. This study may further provide insight into a novel underlying mechanism and a strategy for treating NAFLD.© 2019 John Wiley & Sons, Ltd.

Keyword: fatty liver

Skeletal muscle miR-34a/SIRT1:AMPK axis is activated in experimental and human non-alcoholic steatohepatitis.

Non-alcoholic disease (NAFLD) pathogenesis associates with intramyocellular lipid deposition and mitochondrial dysfunction. microRNAs (miRs), including pro-apoptotic miR-34a, are modulated during disease progression in tissue and plasma. We aimed to investigate the functional role of the miR-34a/SIRT1:AMP-activated protein kinase (AMPK) pathway in modulating local mitochondrial dysfunction in the skeletal muscle of human and experimental non-alcoholic steatohepatitis. Muscle biopsies were obtained from morbid obese NAFLD patients undergoing bariatric surgery. C57BL/6N mice were fed different NAFLD-inducing diets and C2C12 muscle cells incubated with (PA) in the presence or absence of an AMPK activator, or upon miR-34a functional modulation. Several muscle miRNAs, including miR-34a, were found increased with human NAFLD progression. Activation of the miR-34a/SIRT1:AMPK pathway, concomitant with impairment in insulin signalling mediators and deregulation of mitochondrial-shaping proteins, was evident in C2C12 cells incubated with PA, as well as in the skeletal muscle of all three diet-induced NAFLD mice models. Functional studies established the association between miR-34a- and PA-induced muscle cell deregulation. Of note, activation of AMPK almost completely prevented miR-34a- and PA-induced cellular stress. In addition, the miR-34a/SIRT1:AMPK pathway and mitochondrial dynamics dysfunction were also found amplified in muscle of human NAFLD. Finally, muscle miR-34a expression and mitofusin 2 (Mfn2) protein levels correlated with hallmarks of NAFLD and disease progression. Our results indicate that activation of the miR-34a/SIRT1:AMPK pathway leads to mitochondrial dynamics dysfunction in skeletal muscle of human and experimental NAFLD, representing an appealing prospective target in metabolic syndrome. KEY MESSAGES: Skeletal muscle microRNAs are modulated during NAFLD progression. -induced muscle cell dysfunction occurs, at least in part, through activation of the miR-34a/SIRT1:AMPK pathway. miR-34a/SIRT1:AMPK activation associates with mitochondria dynamics dysfunction in human NAFLD.

Keyword: fatty liver

Moderate chronic ethanol consumption exerts beneficial effects on nonalcoholic in mice fed a high-fat diet: possible role of higher formation of triglycerides enriched in monounsaturated acids.

Several clinical studies suggested that light-to-moderate alcohol intake could alleviate nonalcoholic disease (NAFLD), but the underlying mechanism is still poorly understood.Mice fed a high-fat diet (HFD) were submitted or not to moderate ethanol intake for 3\xa0months (ca. 10\xa0g/kg/day) via drinking water. Biochemical, analytical and transcriptomic analyses were performed in serum and .Serum ethanol concentrations in ethanol-treated HFD mice comprised between 0.5 and 0.7\xa0g/l throughout the experiment. NAFLD improvement was observed in ethanol-treated HFD mice as assessed by reduced serum transaminase activity. This was associated with less microvesicular and more macrovacuolar steatosis, the absence of apoptotic hepatocytes and a trend towards less fibrosis. lipid analysis showed increased amounts of acids incorporated in triglycerides and phospholipids, reduced proportion of in total lipids and higher desaturation index, thus suggesting enhanced stearoyl-coenzyme A desaturase activity. mRNA expression of several glycolytic and lipogenic enzymes was upregulated. Genome-wide expression profiling and gene set enrichment analysis revealed an overall downregulation of the expression of genes involved in collagen fibril organization and leukocyte chemotaxis and an overall upregulation of the expression of genes involved in oxidative phosphorylation and mitochondrial respiratory chain complex assembly. In addition, mRNA expression of several proteasome subunits was upregulated in ethanol-treated HFD mice.Moderate chronic ethanol consumption may alleviate NAFLD by several mechanisms including the generation of non-toxic lipid species, reduced expression of profibrotic and proinflammatory genes, restoration of mitochondrial function and possible stimulation of proteasome activity.

Keyword: fatty liver

-Enriched Diet Induces Hepatic Steatosis and Injury in Adult Zebrafish.

(PA) is the most abundant saturated in fast foods and is known to induce inflammation and cellular injury in various tissues. In this study, we investigated whether a PA-enriched diet can induce hepatic steatosis and injury in adult zebrafish. The adult zebrafish exhibited increased body weight, hyperlipidemia, hyperglycemia, and steatosis and a hepatic injury phenotype after being fed with a PA-enriched diet for 6 weeks. The quantitative polymerase chain reaction analysis demonstrated that genes associated with hepatic injury were all significantly increased in the . Furthermore, livers from the PA-fed group showed an increased messenger RNA (mRNA) expression associated with oxidative stress and endoplasmic reticulum (ER) stress responses. We also found significant upregulation of genes involved in lipid metabolism and triacylglyceride accumulation. Ultrastructural analysis revealed mitochondrial cristae injury and a dilated ER phenotype in the PA-fed hepatocytes, which can be causes of hepatic injury. PA-enriched diet induced hepatic steatosis and injury in adult zebrafish that recapitulated typical metabolic changes and pathophysiological changes as well as increased oxidative stress and ER stress observed in patients with nonalcoholic disease.

Keyword: fatty liver

Apigenin reduces the excessive accumulation of lipids induced by via the AMPK signaling pathway in HepG2 cells.

In recent years, increasing attention has been paid to diseases caused by excessive accumulation of lipids in the with therapeutic agents derived from natural products offering an alternative treatment to conventional therapies. Among these therapeutic agents, apigenin, a natural flavonoid, has been proven to exert various beneficial biological effects. In the present study, the antiadipogenic effects of apigenin in HepG2 cells was investigated. It was demonstrated that the treatment of cells with different concentrations of apigenin for 24 h significantly decreased the -induced increases in total cholesterol (TC) and triglyceride (TG) levels as well as intracellular lipid accumulation. In addition, apigenin increased the phosphorylated-AMP-activated protein kinase (AMPK) levels but decreased the expression levels of 3-hydroxy-3-methylglutaryl CoA reductase, sterol regulatory element-binding protein (SREBP)-1, synthase, and SREBP-2 in a concentration-dependent manner. The present findings suggested that apigenin might improve lipid metabolism by activating the AMPK/SREBP pathway to reduce lipid accumulation in the .Copyright: © Lu et al.

Keyword: fatty liver

Role of caveolin-1 in hepatocellular carcinoma arising from non-alcoholic disease.

The molecular features of hepatocellular carcinoma arising from non-alcoholic disease (NAFLD-HCC) are not well known. In this study, we investigated the mechanism by which NAFLD-HCC survives in a fat-rich environment. We found that caveolin (CAV)-1 was overexpressed in clinical specimens from NAFLD-HCC patients. HepG2, HLE, and HuH-7 HCC cell lines showed decreased proliferation in the presence of the saturated acids and stearic , although only HLE cells expressed high levels of CAV-1. HLE cells treated with oleic (OA) showed robust proliferation, whereas CAV-null HepG2 cells showed reduced proliferation and increased apoptosis. CAV-1 knockdown in HLE cells attenuated the OA-induced increase in proliferation and enhanced apoptosis. Liquid chromatography-tandem mass spectrometry analysis revealed that the levels of OA-containing ceramide, a pro-apoptotic factor, were higher in HepG2 and CAV-1-deficient HLE cells than in HLE cells, suggesting that CAV-1 inhibits apoptosis by decreasing the level of OA-containing ceramide. These results indicate that CAV-1 is important for NAFLD-HCC survival in -rich environments and is a potential therapeutic target.© 2018 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

Keyword: fatty liver

Ground Beef High in Total Fat and Saturated Acids Decreases X Receptor Signaling Targets in Peripheral Blood Mononuclear Cells of Men and Women.

We hypothesized that consumption of saturated acids in the form of high-fat ground beef for 5 weeks would depress X receptor signaling targets in peripheral blood mononuclear cells (PBMC) and that changes in gene expression would be associated with the corresponding changes in lipoprotein cholesterol (C) concentrations. Older men (n = 5, age 68.0 ± 4.6 years) and postmenopausal women (n = 7, age 60.9 ± 3.1 years) were assigned randomly to consume ground-beef containing 18% total fat (18F) or 25% total fat (25F), five patties per week for 5 weeks with an intervening 4-week washout period. The 25F and 18F ground-beef increased (p < 0.05) the intake of saturated fat, monounsaturated fat, , and stearic , but the 25F ground-beef increased only the intake of oleic (p < 0.05). The ground-beefs 18F and 25F increased the plasma concentration of (p < 0.05) and decreased the plasma concentrations of arachidonic, eicosapentaenoic, and docosahexaenic acids (p < 0.05). The interventions of 18F and 25F ground-beef decreased very low-density lipoprotein C concentrations and increased particle diameters and low-density lipoprotein (LDL)-I-C and LDL-II-C concentrations (p < 0.05). The ground-beef 25F decreased PBMC mRNA levels for the adenosine triphosphate (ATP) binding cassette A, ATP binding cassette G1, sterol regulatory element binding protein-1, and LDL receptor (LDLR) (p < 0.05). The ground-beef 18F increased mRNA levels for stearoyl-CoA desaturase-1 (p < 0.05). We conclude that the increased LDL particle size and LDL-I-C and LDL-II-C concentrations following the 25F ground-beef intervention may have been caused by decreased hepatic LDLR gene expression.© 2018 AOCS.

Keyword: fatty liver